WO2006122476A1

WO2006122476A1 - A method of inactivating virus, treatment system and device used thereof

Info

Publication number: WO2006122476A1
Application number: PCT/CN2006/000877
Authority: WO
Inventors: Fanglin Zou; Chunsheng Chen; Jianxia Wang
Original assignee: Chengdu Kuachang Medical Industrial Limited
Priority date: 2005-04-29
Filing date: 2006-04-29
Publication date: 2006-11-23

Abstract

A method of inactivating virus, at least comprising: A) providing bio-fluid; B) providing the treatment system; and C) contacting the bio-fluid with the treatment system, wherein the treatment system at least contains solid-phase medium and the chambers containing the solid-phase medium, which at least comprises: a) the adsorptive medium of the viral inactivator; and /or b) the viral inactivating medium, which comprises the viral inactivator and the adsorptive medium of the viral inactivator, which comprises: (a) deactivation adsorptive medium, which at least contains the adsorptive medium and the deactivator fixed on the surface of the medium; and/or (b) physical adsorptive medium, which comprises photosensitizer physically adsorptive medium as photosensitizer viral inactivator’s adsorptive medium, and organic solvent physically adsorptive medium as organic solvent viral inactivator’s adsorptive medium.

Description

Virus inactivation method, processing system and device used therein

The present invention relates generally to methods of treating biological fluids, and more particularly to virus inactivation, treatment systems, and devices.

Background technique

Since the 1970s, incidents of serious medical health consequences due to contamination of pathogens in biological materials, such as viruses (vims), have become a growing concern of the public. To this end, the scientific and technological community has developed several techniques for inactivating or/or removing these pathogens. In the following, the virus inactivation, especially the virus-killing activity containing the solid-liquid phase reaction, is taken as an example to illustrate the problems to be solved in the prior art.

The existing virus inactivation technique containing solid-liquid phase reaction is mainly based on organic solvent virus inactivation (Solvant-Detergent treatment, referred to as SD treatment in the present invention). The organic solvent virus inactivating medium contains an organic solvent S (which may additionally contain detergent D) and a virus inactivating agent adsorbent such as activated carbon (refer to Chinese Patent 01123658.2). Other virus inactivation techniques containing solid-liquid phase reactions, including immobilization of alkylphosphorus compounds (using inorganic adsorbents as adsorbents, reference to German patent DE40 (H099A1), immobilized iodine treatment (with silicon-containing dry soil PVPP) The filter plate is used as an adsorbent, refer to the product introduction of the German Shenk company, etc. The problem of the prior art is that the activated carbon, the inorganic adsorbent or the silicon-containing dry soil PVPP filter plate has side effects (for example, strong protein adsorption capacity, Inactivation of coagulation factors, a significant increase in plasma APTT values, etc.).

In the prior art in which a solid phase medium is adsorbed to remove an unwanted virus inactivating agent, the organic solvent virus inactivating agent adsorption medium is an inorganic adsorbent (for example, a perlite filter plate containing perlite:), an affinity adsorbent (for example). C-18 oleophilic resin), activated carbon. A problem with the prior art is that the affinity adsorbent is expensive and not suitable for single use, and the activated carbon and inorganic adsorbent used have side effects (e.g., strong protein adsorption capacity, inactivation of coagulation factors, and substantial increase in plasma APTT value, and many more). ,

In the prior art in which a solid phase medium is adsorbed to remove an unwanted toxic inactivating agent, the photosensitizer virus inactivating agent adsorption medium is activated carbon (for example, PCT/US94/U227; US6159375; US08/347564), and Ion exchange resin (WO9103933), poly styrene-divinylbenzene, acrylic ester polymers (US6348309). A problem with the prior art is that activated carbon, silica, and ion exchange resins have side effects (e.g., strong protein adsorption capacity, a large increase in plasma APTT value, etc.), while macroporous adsorption resins tend to fall off and contaminate biological fluids. Side effects can also be formed.

In summary, there are existing methods for treating biological fluids, including virus inactivation treatments, where Solid phase media in the system have many side effects, such as: protein conformational change and amount loss; platelet conformational change and amount loss; red blood cell conformational change and amount loss, introduction of impurity particles, and the like. A similar phenomenon exists in the existing leukocyte removal technology. Thus, under current conditions, the development of a virus inactivation method that has both effective functions (eg, adsorption of virus inactivating agents and/or virus inactivation) with minimal side effects is still the pursuit of science and technology. The goal.

Summary of the invention

It is an object of the present invention to provide a virus inactivation method that has both an effective virus inactivating function, or/and an effective viral inactivating agent removal function, with substantially no or only minimal side effects, and is useful in the present invention. Method of processing systems and devices. In the present invention, the expression "effective virus inactivating function" means

More than 99.9% of the functions of one or more model viruses are inactivated; the expression "effective virus inactivating agent removal function" means a function of removing more than 90% of one or more virus inactivating agents.

The basic technical solution of the method of the present invention is to achieve the object of the present invention by using a solid phase medium having both effective functions and only minimal side effects. The technical solution includes an embodiment: 1. In general, the addition of a oximation agent to a solid phase medium minimizes side effects in virus inactivation; 2. For a particular virus inactivating agent, a selection function/ Side effects can be achieved for the same purpose as the maximum solid phase medium.

The virus inactivation method of the present invention BJ, comprising at least: Α) providing a biological fluid; Β) providing a treatment system; and C) contacting the biological fluid with the treatment system, wherein the treatment system comprises at least a solid a phase medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, the virus inactivating medium comprising a virus An inactivating agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: (a) a passivation adsorption medium, the passivation adsorption medium containing at least an adsorption medium and being fixed in adsorption a passivating agent on the medium; or/and (b) a physical adsorption medium, the physical adsorption medium comprising: (i) a photosensitizer physical adsorbing fiber used as a photosensitizer virus inactivating agent adsorption medium, the photosensitizer Physically adsorbing fibers containing an organic polymer physical adsorbent of a photosensitizer virus inactivating agent; and (ii) an organic solvent physical adsorption medium used as an organic solvent virus inactivating agent adsorption medium, the organic solvent physical adsorption medium containing an organic solvent The organic polymer physical adsorbent drug inactivating agent.

The treatment system of the present invention is the treatment system described in the above virus inactivation method of the present invention. The processing system comprises at least a solid phase medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, The virus inactivating medium comprises a virus inactivating agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: (a) a passivation adsorption medium, the passivation adsorption medium being at least a passivating agent comprising an adsorbent medium and immobilized on the adsorbent medium; or/and (b) a physical adsorbent medium comprising: (i) a photosensitizer physics used as a photosensitizer virus inactivating agent adsorption medium Adsorbing fibers, the photosensitizer physically adsorbing an organic polymer physical adsorbent containing a photosensitizer virus inactivating agent; and (ii) using as an organic solvent virus inactivating agent An organic solvent physical adsorption medium of the medium, the organic solvent physical adsorption medium containing an organic solvent physical adsorbent of the organic solvent virus inactivating agent.

The device of the present invention comprises at least the above described processing system.

Embodiments of the invention will illustrate that the objects of the invention can be attained by the method, processing system or apparatus of the invention. It is to be noted that the method, treatment system or device of the present invention is particularly suitable for the case where the solid phase medium is used at one time, in addition to the advantages of its small side effects.

detailed description

It should be understood by those skilled in the art that the terms used below are merely used to describe the specific modes of the present invention, and the present invention may not be limited to the definitions explained below.

In the present invention, the term "biological liquid" means a liquid containing or possibly containing a biological substance such as blood and blood components, plasma and plasma derivatives, biological products, biopharmaceuticals, and the like. In the present invention, the term "single human blood component" means blood collected from one or more times from the same person or/and components separated therefrom, including single blood, single plasma, single serving. Platelets, single red blood cells, and more.

In the present invention, the term "effectively reducing pathogen harm" means a treatment process including effective virus inactivation or/and leukocyte removal; the term "viral inactivation" means by adding a virus inactivating agent (for example)

SD agent, photosensitizer virus inactivating agent, etc.) or/and physical energy (eg heat, light, etc.), a process for effectively reducing the infectious activity of at least a portion of the virus present in the preparation; the term "white blood cell removal" "Includes a process for reducing leukocyte concentration by mechanisms such as adsorption and size filtration of leukocytes; the expression "effective" means, for virus inactivation, 99.9% or more of one or more model viruses are inactivated, For leukocyte removal, it means that more than 90% of white blood cells are removed.

In the present invention, the term "viral inactivation treatment" refers to the entire process of bringing a virus inactivating agent into contact with a biological fluid until the viral inactivating agent in the biological fluid is substantially removed. Viruses include membrane viruses or/and non-lipid envelope viruses. The virus inactivation treatment contains a virus inactivation process, and in many cases, a virus inactivating agent removal process. In the production process, the virus inactivation treatment and other treatments are performed sequentially or/and simultaneously, and thus the above virus inactivation treatment may also partially undergo or/and be accompanied by other treatments other than virus inactivation (eg, protein) Purification, etc.).

In the present invention, the term "viral inactivating agent" means any additive which has a virus inactivating function for all or part of viruses which may be present in a biological fluid, but may not be limited to a virus inactivating function, such as a photosensitizer virus inactivating agent/ Photosensitizer virus inactivating agent in light treatment, organic solvent virus inactivating agent in SD treatment or organic solvent virus inactivating agent/detergent, and the like. In many cases, the virus inactivating agent may have other functions besides the virus inactivating function. For example, psoralen is inactivated by viruses, but also inactivated by some other organisms (such as bacteria), in which case the virus inactivating agent is a bacterial inactivating agent, and the like. The term "viral inactivation agent" "Biological" is a derivative of a virus inactivating agent formed in a virus inactivation treatment, such as a photosensitizer virus inactivating agent/a derivative of methylene blue which is a virus inactivating agent during photoactivation (for example, azure) A and B).

In the present invention, the organic solvent virus inactivation comprises virus inactivation by a combination of an organic solvent virus inactivating agent (S) or an organic solvent virus inactivating agent and a detergent combination (SD) for the virus inactivating agent. . The SD treatment includes an SD treatment method invented by the New York Blood Center (US Pat. No. 4,540,573), and a derivative method developed by others (for example, a virus-in addition method in which only S is added, and a virus inactivation treatment method in which S is shared with alcohol, etc.). In the present invention, the term "organic solvent virus inactivating agent (abbreviated as S in the present invention)" means an organic solvent used under specific conditions of virus inactivation treatment. These specific conditions include, but are not limited to: competitive adsorption (eg, in biological fluid environments often contain components that affect the adsorption of photosensitizers, such as lipids in human plasma); reaction kinetic conditions are less favorable (eg, liquid phase) The concentration of the virus inactivating agent when the virus is inactivated is generally not more than 1% (w/v) _{; the} reaction conditions are relatively conservative (for example, the reaction temperature is usually less than 38 ° C and the pH is between 4 and 10); Therefore, from the viewpoint of the technical solution, the organic solvent virus inactivating agent in the present invention is distinguished from the organic solvent in the general sense. The organic solvent virus inactivating agent includes any solvent for virus inactivation, such as tributyl phosphate (abbreviated as TnBP in the present invention), formaldehyde, diethyl ether, and the like. It is worth noting that organic solvent virus inactivating agents are often shared with detergents. In the present invention, the detergent includes any detergent for virus inactivation, such as a Tween-type detergent (e.g., Tween-80) and a Triton-based detergent (e.g., Triton X 100).

In the present invention, the photosensitizer virus inactivation comprises virus inactivation with a photosensitizer virus inactivating agent as a virus inactivating agent; the term "photosensitizer virus inactivating agent" means under specific conditions of virus inactivation treatment. The photosensitizer used. These specific conditions include, but are not limited to: competitive adsorption (eg, in biological fluid environments often contain components that affect the adsorption of photosensitizers, such as lipids in human blood plasma); reaction kinetic conditions are less favorable (eg, liquid The concentration of the virus inactivating agent when the phase virus is inactivated is generally not more than Ιμπιοΐ/l); the reaction conditions are relatively conservative (for example, usually the reaction temperature is less than 38*^ and 11 is between 6-10); Thus, from the viewpoint of the technical solution, the photosensitizer virus inactivating agent in the present invention is distinguished from the photosensitizer in the general sense. Typically, photosensitizer virus inactivating agents include, but are not limited to, dye photosensitizer virus inactivating agents, hypericin and psoralen photosensitizer virus inactivating agents, and the like. Generally, these photosensitizer virus inactivating agents can have much higher virus inactivation efficiencies under appropriate light exposure. In the present invention, the term "dye-based photosensitizer virus inactivating agent" means a dye-like substance which can be used as a photosensitizer virus inactivating agent for virus inactivation (for example, photochemical virus inactivation), for example, phthalocyanine (US Patent No. 5,232,844) ), sapphyrin (US Patent US5041078), phenothiazine dyes (Lambrecht et al., Vox Sang. 60, 207-213, (1991), and cyanine dyes (US Patent US 4,915,683), etc.; "Psoralen-type photosensitizer virus inactivating agent" means that the photosensitizer virus inactivating agent can be used for virus inactivation (eg photochemistry) Viral inactivated psoralen substances (U.S. Patents 4,169,204, 4,294,822, 4,328,239 and 4,727,027), including psoralen (English Psoralen^ psoralen derivatives, etc. Examples of psoralen derivatives) There are 8-methoxyproralen 5-methoxy psoralen S59 psoralen^ trioxsalen-, aminomethyltrimethylpsorale and so on.

In the present invention, the term "side effects" refers to the ability of a solid phase medium, treatment system or device to have unwanted functions in addition to having a useful function, and may also cause an undesirable change in the biological activity of the biological fluid, such as the following One or more changes: a reduction in the total amount of protein, interference with the coagulation system, changes in platelet morphology, improvement in hemolytic properties, and the like. In the present invention, the term "specific adsorption" refers to adsorption of a virus inactivating agent to a virus inactivating agent, or removal of leukocyte media by leukocyte medium; the term "non-specific adsorption" refers to a virus inactivating agent adsorption medium pair Adsorption of substances other than viral inactivating agents, or removal of substances other than leukocytes by leukocyte mediators. Non-specific adsorption includes adsorption (e.g., degreasing) and unwanted adsorption (e.g., reduction in clotting factor activity) that are beneficial to the desired properties of the biological fluid. Unwanted reactivity of the solid phase medium includes unwanted adsorption and unwanted other reactive activities (e.g., catalytic activity, enzyme activating activity, activity involved in reactions in biological fluids, etc.).

In the present invention, the term "solid phase medium", also referred to as medium, refers to a solid phase material having a certain function (for example, adsorption function, virus inactivation function, etc.); the term "virus inactivating agent adsorption medium", Sometimes referred to as an adsorption medium, it means having at least, but not limited to, a function of adsorbing a virus inactivating agent or/and a virus inactivating agent derivative (for example, sometimes it may also have a filtering function, a leukocyte function, etc.), And can ultimately reduce the concentration of viral inactivating agents or/and virus inactivating agent derivatives in biological fluids to acceptable levels (eg, tributyl phosphate concentration less than 10 ppm, methylene blue concentration less than 0.03 _g / ml, etc. The solid phase material, the adsorption medium may be a pure substance (such as activated carbon powder, wood fiber, etc.), or may be a mixture (for example, a cellulose-containing filter medium, an activated carbon-containing filter medium, a C18 chromatography gel, etc.) The term "viral inactivating medium" means a solid phase medium having at least, but not necessarily limited to, a virus inactivating function (for example, it may also have a filtering function sometimes), such as a virus inactivating agent/adsorption medium complex, the term" Interleukin "refers to any solid phase media may be used to leukocytes.

In the present invention, the term "viral inactivating agent adsorbent" means an adsorbent which is effective for removing a virus inactivating agent by adsorption under specific conditions of virus inactivation treatment. These specific conditions include: specific viral inactivating agents; specific adsorption environments (eg, biological fluids often contain components that affect adsorption, such as lipids in human plasma); specific adsorption kinetic conditions (eg, eg In the embodiment of the present invention, the concentration of the virus inactivating agent in the biological liquid is small, but the concentration of the virus inactivating agent remaining after the adsorption is required to be extremely low, for example, less than 10 ppm or less than 0.01 μηιο 1 / 1) _; specific adsorption reaction conditions ( For example, to maintain the activity of the biological fluid, the adsorption temperature is usually limited to 0-37 ° C, the adsorption pH is usually limited to 3.8-9.8); Thus, from the viewpoint of the technical solution, the virus inactivating agent adsorbent in the present invention is distinguished from the adsorbent in the general sense. In the present invention, the term "organic polymer chemical adsorbent" is prepared by introducing a sufficient amount of a bonding group (for example, an ionic group, a coordinating group, etc.) which generates a chemical bond to an organic polymer, thereby Mainly through the introduced bonding groups to generate chemical bonds (such as ionic bonds, coordination bonds, etc.) into the reagents, such as ion exchange fibers, ion exchange particles and chelating agents; "organic 髙 molecular affinity adsorption "agent" is prepared by introducing a sufficient amount of an affinity ligand (eg, a terminal group of hydrazine, or an affinity of a viral RNA or DNA for the affinity adsorption of certain photosensitizer virus inactivating agents) onto an organic hydrazine molecule, Therefore, the reagent mainly adsorbed by the introduced affinity ligand, such as affinity fiber, affinity particle, etc.; "organic polymer physical adsorbent" refers to the above organic polymer chemical adsorbent and organic polymer affinity Organic polymeric adsorbents other than adsorbents, such as plant fibers, polyphenylenes, and the like, include organic polymers that have not undergone specific affinity grouping or/and have not undergone special ion exchange group modification. . It should be noted that the organic polymer does not include activated carbon. In the embodiment of the present invention, the organic polymer physical adsorbent does not include the currently known organic polymer affinity adsorbent and the organic polymer ion exchange adsorbent, and these functionalized organic polymers are subjected to special affinity grouping ( Modifications such as affinity fibers, affinity particles) or/and specialized ion exchange groups (eg, ion exchange fibers, ion exchange particles). The organic polymer physical adsorbent does not have to be prepared by introducing a sufficient amount of ion exchange groups or organic ligands onto the organic polymer, and thus mainly through weak interactions on the organic polymer (including van der Waals attraction, dipole-dipole Interaction, hydrogenation, etc.) and adsorption of a small amount of (end group, or / and 5% or less substituents on the chain) ionic groups (for example, a small amount of acid groups on plant fibers) introduced during the preparation process Reagents.

In the present invention, the organic polymer in the term "organic polymer physical adsorbent" includes a natural polymer, such as a strong natural polysaccharide. In an embodiment of the invention, the polysaccharide comprises one or more of the following: cellulose, dextran, agarose, chitosan, starch. Examples of the cellulose derivative include nitrocellulose, cellulose acetate, methyl cellulose, carboxymethyl cellulose, and the like. Although polyglycans have been used as carriers in addition to viral inactivating agents, for example, nitrocellulose has been used as a carrier for photosensitizer virus inactivating agent ligands in addition to photosensitizer virus inactivating agents and leukocyte-removing filters (US patent) US08/179567, 08/204102, 08/347564), a polysaccharide containing polypolysaccharide is used as a carrier for an SD agent adsorbent (for example, C18) for use in addition to an SD agent, etc., another meaning of the embodiment of the present invention Unexpected results are a sufficiently strong, kinetically stable, reproducible adsorption of the polysaccharide itself to the dye (e.g., methylene blue). In an embodiment of the invention, certain filters (e.g., Seitz-BiolO, Seitz-Bio20, Seitz-bio 40), certain porous particles (e.g., Sephadex G10, Sephadex G25 Sephadex G50, Sepharose) all contain a polysaccharide.

In the present invention, the organic polymer in the term "organic polymer physical adsorbent" includes synthetic polymers such as polyolefins, polyurethanes, polydecyl esters, and the like. In the embodiment of the present invention, the synthetic polymer includes a polyolefin such as polystyrene, polyvinyl chloride, polyvinyl alcohol, or the like. In an embodiment of the invention, certain filters (eg Seitz-Supradurl00, Seitz-Supradur200, Seitz-Supradur 500), certain Porous particles (for example, crosslinked acrylic macroporous adsorption resin, crosslinked polystyrene macroporous adsorption resin, polystyrene-polyacrylonitrile macroporous resin, polymethyl methacrylate resin, polyisobutylene, etc.) all contain poly Olefins.

In the present invention, the term "passivating agent" refers to a reagent which has a passivation effect on an undesired reaction (eg, side effects) on a solid phase medium or other composition of a treatment system, but may not be limited to passivation. In the present invention, the term "composite" means a material in which two or more compositions are joined by adsorption or other physicochemical, chemical, or biochemical interaction. For example, the term "passivation agent/adsorption medium complex" "refers to a complex of at least an adsorbent medium composed of a virus inactivating agent and a passivating agent of an adsorbent medium; the term "viral inactivating agent / adsorbent medium / passivator complex" means a minimum composition of a virus inactivating agent a complex of an adsorption medium for the virus inactivating agent and a passivating agent for the adsorption medium.

In the present invention, the processing system can be used as a device (eg, a filter, a chromatography column, etc.), or as a component of the device (eg, a filter, column, etc. in a single blood or component processing device). Wait). In the present invention, the term "device" means a structure in which the minimum composition is the above-described treatment system, which can be practically applied to the treatment of biological fluids, such as filters, chromatography columns, kits (English Kit:), etc.; "Case" means a hollow vessel that can hold at least a solid phase medium, such as a hollow filter cartridge of a filter, a hollow column of a chromatography column, and the like.

In the present invention, the term "filter material" generally means a solid phase medium which can only allow a structure below a certain size to flow. Therefore, the filter medium in the present invention is often used as a stationary phase in a solid-liquid phase reaction, for example, a filter material having an adsorption function for a virus inactivating agent (referred to as a specific adsorption filter medium). Whether a filter material specifically adsorbs the filter material, in addition to the information provided by the manufacturer (such as activated carbon filter plate), it is mainly to see if it has a specific adsorption function in the virus inactivation treatment. For example, a Seitz-Bio filter plate which is generally considered to have no adsorption function is found to be a dye-based photosensitizer virus inactivating agent adsorption medium in the examples of the present invention. The filter media includes absolute filter media and depth filter media. A preferred embodiment of the filter material of the present invention is a depth filtration filter. Although not intended to be discussed in theory, the fluid flow path in the deep filter media is long and has a significant impact on its function (eg adsorption, virus inactivation). Filter media come in many forms, such as filters, filter plates, filter rods, filter cartridges, filter mats, and more.

In the present invention, the term "filter" means a device comprising at least a filter material and its holder, including but not limited to known filters. The holder includes a structure that seals the periphery of the solid phase medium with pressure, such as a filter disc, a filter cartridge, a filter column, and the like. In the filter of the following embodiments of the present invention, the holder generally includes: a container for accommodating the solid phase medium, an upper end structure and a lower end structure for fixing the solid phase medium, and a pressure sealing filter for surrounding the solid phase medium The structure of the solid-state medium of the material (for example, a pressure-closed structure in which the solid phase medium is closed to avoid liquid leakage due to the contact force of the convex body which is higher on the upper end structure and the lower end structure to the periphery of the solid phase medium when closed). In addition, other functional structures can be added, such as in the upper structure. Or a structure with a special function introduced on the lower end structure (for example, a sterile filter membrane, etc.). Those skilled in the art will appreciate that a plurality of filters having different uses can be prepared by using a filter device of the following embodiments of the present invention by combining with different structures.

In the present invention, the term "dyeing" means a structure which can bind a dye, including a physical structure, a physicochemical structure, a chemical structure (for example, a group), and the like; the term "fiber" means a length/diameter ratio of more than 10, and A material having an average diameter of less than 1 mm; the term "Fibrets" refers to a fiber having a large number of irregular micro branches and a large surface area formed by a special production process or special processing. Structure, this fiber microstructure is very short (less than 1 mm) and very fine (less than 50 μm), for example Seitz-BiolO, Seitz-Bio20, Seitz-bio40 filter plates contain fiber microstructure.

A first aspect of the invention relates to a virus inactivation method of the invention.

The virus inactivating method of the present invention comprises at least: A) providing a biological fluid; B) providing a treatment system; and C) contacting the biological fluid with the treatment system, wherein the treatment system comprises at least a solid phase a medium and a chamber containing the solid phase medium, wherein the solid phase medium comprises at least: a) a virus inactivating agent adsorption medium; and/or b) a virus inactivating medium, the virus inactivating medium comprising a virus a living agent adsorption medium and a virus inactivating agent immobilized thereon, wherein the virus inactivating agent adsorption medium comprises: 0) a passivation adsorption medium, the passivation adsorption medium containing at least an adsorption medium and being fixed on the adsorption medium a passivating agent; or/and (b) a physical adsorbing medium comprising: (i) a photosensitizer physic adsorbing fiber used as a photosensitizer virus inactivating agent adsorption medium, said photosensitizer physically adsorbing An organic hydrazine molecular physical adsorbent containing a photosensitizer virus inactivating agent; and (ii) an organic solvent physical adsorption medium used as an organic solvent virus inactivating agent adsorption medium, the organic solvent physical adsorption medium containing organic solvent The organic polymer physical adsorbent inactivating agent.

The method of the present invention is based on unexpected results in embodiments of the invention:

1). Application for purification of adsorption media:

Since the permissible residual value of the virus inactivating agent in the bio-liquid product standard is usually very low (for example, the TnBP approved residue value in the European Pharmacopoeia is less than 10 ppm), the general view is: It is not suitable for passivation of the adsorption medium. Surprisingly, in some embodiments of the present invention, after the adsorption medium is fixed to form the adsorption medium/passivator compound, the adsorption medium can still be used for adsorption removal of the virus inactivating agent, and has less side effects than the adsorption medium.

In addition, due to the high requirement for virus inactivation (usually requiring a virus titer to drop by more than 10 ³ ), the general view is that the virus is inactivated medium (especially the solid phase of the virus inactivating agent not on the long-chain adsorption group) Passivation of a medium, such as SD-activated carbon, may reduce the inactivation efficiency of the virus inactivating medium. Thus, no passivation treatment was introduced in the development of the virus inactivating medium. However, embodiments of the present invention will demonstrate that the use of a virus inactivating medium containing a passivating agent not only effectively protects the biological activity of the liquid medium contacted by the solid phase medium, but also effectively inactivates the virus, thereby achieving The object of the invention. In addition, the leukocyte-removing medium usually has more or less adsorption capacity to white blood cells. The general idea is that the addition of a passivating agent can inhibit its side effects, but it may also reduce its adsorption on white blood cells. Surprisingly, however, in one embodiment of the invention, the addition of a passivating agent inhibits its side effects, but it can still be used to effectively remove leukocytes. In a preferred embodiment, the first treatment system of the invention can be used to simultaneously remove viral inactivating agents (or/and viral inactivating agent derivatives) and white blood cells.

2). About the application of physical adsorption media:

In general, the removal of additives by solid phase media containing adsorbents is a well-known technique that has long been used. Those skilled in the art will appreciate that the viral inactivating agent can be removed by a solid phase medium containing a viral inactivating agent adsorbent. Since the solid phase medium with adsorption function is generally very much, and the virus inactivating agent includes many kinds, the virus inactivating agent adsorption medium containing the adsorption function for the specific virus inactivating agent is developed. Certain processing systems have been a very difficult creative task, and it has become more difficult to develop a treatment system that has a specific adsorption of a specific viral inactivating agent without substantial non-specific adsorption (for example, essentially no side effects:). Creative work.

The use of photosensitizers for physically adsorbing fibers is also based on the unexpected results of embodiments of the present invention. For example, although: A) certain dyes (eg, methylene blue) are one of the most commonly used viral inactivating agents in human blood or human blood components; B) fibers containing dyed seats (eg, natural fibers) can adsorb dyes; C) Even filter media containing natural fibers have long been used for the production of biological products, but no adsorption medium for directly using natural fibers as a dye-based photosensitizer virus inactivating agent has been seen so far. This may be familiar, whether it is capable of satisfying the dye-based photosensitizer virus inactivating agent at a lower concentration (usually less than 1μπι ₀ 1/ιη1) and a low temperature (usually less than 36Ό). Effective adsorption lacks confidence. However, according to an embodiment of the present invention, it is surprising that the use of synthetic fibers of natural fibers or non-ion exchange fibers for the effective removal of dye virus inactivating agents is no more difficult and even more difficult than dyeing fibers. Easier to implement. For example, methylene blue is generally not used for dyeing natural fibers, but natural fibers are used to effectively remove methylene blue virus inactivating agents. According to an embodiment of the present invention, these fibers are not only adsorbable to the dye-based photosensitizer virus inactivating agent, but also have the characteristic that the adsorption is sufficiently strong (for example, the concentration of methylene blue after adsorption is reduced to 0.03 g / Mol is below); its adsorption is kinetically stable (for example, there is no significant desorption of methylene blue adsorbed on the filter plate when the biological liquid is continuously flowing); its adsorption is reproducible (for example, under the same conditions) The adsorption reproducibility is 100%); and its adsorption is sufficiently specific (for example, substantially no side effects).

The present invention is not intended to be discussed in theory, and only some of the fibers containing organic polymeric physical adsorbents are provided for some observations. Perhaps due to the following reasons, weak interactions of organic polymeric physical adsorbents (such as trace ionic groups introduced during production, van der Waals attraction, dipole-dipole interaction, hydrogen chemistry) are strong enough to effectively adsorb dyes Photosensitizer virus inactivating agent: 1). Dye-based photosensitizer The "dye up-take" of the virus inactivating agent is usually a biological liquid containing a fiber as a stationary phase and a dye-based photosensitizer virus inactivating agent. Flow The phase is carried out under the conditions of the preferred solid-liquid phase reaction; 2) the dye-based photosensitizer virus inactivating agent is not required to be uniformly dyed; 3) the dye-based photosensitizer virus to be adsorbed in terms of the amount of fibers used The total amount of inactivating agent is not very large; ₄ ). The structural complexity of the fiber; In summary, these weak interactions of organic polymeric physical adsorbents are sufficiently strong for specific viral inactivating agents under specific conditions, and the side effects of biological fluids under specific conditions are small enough. It has a relatively high specific adsorption capacity, so that the object of the present invention can be achieved.

The use of organic solvent physical adsorption media is also based on the unexpected results of embodiments of the present invention. For example, although: A) an organic solvent virus inactivating agent (eg TnBP) is one of the most commonly used viral inactivating agents in human blood or human blood components; B) porous particles containing polyolefin can remove oil stains; C) even containing Porous particles of polyolefin have been used for the removal of dye-type virus inactivating agents, however, no porous particles containing polyolefin have been used as an adsorption medium for organic solvent virus inactivating agents, especially for organic solvent viruses. The inactivating agent is combined to prepare a virus inactivating medium. Similar to the photosensitizer virus inactivating agent described above, this may be familiar, perhaps because its adsorption capacity can be met at lower concentrations (usually less than 1% w/v) and at low temperatures (usually less than 37°). Under the conditions of C), the effective adsorption of the organic solvent virus inactivating agent lacks confidence. However, according to an embodiment of the present invention, the porous particles not only have an adsorption capacity to the organic solvent virus inactivating agent, but also have the characteristic that: the adsorption is sufficiently strong (for example, the concentration of TnBP after adsorption is reduced to less than 10 ppm); The adsorption is kinetically stable (eg, no significant TnBP desorption occurs); its adsorption is reproducible (eg, its adsorption reproducibility is 100% under the same conditions); and its adsorption is sufficiently specific (eg , basically no side effects).

In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises an organic solvent virus inactivating agent adsorption medium, and the organic solvent virus inactivating agent adsorption medium contains at least the passivation adsorption medium. Or / and organic solvent physical adsorption medium.

In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises an organic solvent virus inactivating medium, and the organic solvent virus inactivating medium contains at least an organic solvent virus inactivating agent, and the blunt The adsorption medium, or/and the organic solvent physical adsorption medium.

In an embodiment of the virus inactivation method of the present invention, the organic solvent physical adsorption medium comprises a macroporous adsorption resin or/and a fiber. In the embodiment of the present invention, the fiber (organic solvent physical adsorption fiber) contains an organic polymer physical adsorbent containing an organic solvent virus inactivating agent, such as natural fiber (for example, Seitz Bio series filter plate), polyolefin fiber (for example, Seitz). Supradur 80, Seitz Supradur500, Seitz Eco 1000, etc.), etc. The macroporous adsorption resin (organic solvent physical adsorption resin) used includes polystyrene particles (for example, Amberlite XAD-7HP, Amberlite AD-16), phenolic polycondensation type particles, polyacrylate particles, and polyalkyl ester-containing macroporous adsorption resin. In addition to the above, the organic solvent physical adsorption medium further includes a porous non-particulate material having an inner surface containing the organic polymer. For example, polyolefin coated ceramics, and many more.

In an embodiment of the virus inactivation method of the present invention, the organic polymer physical adsorbent of the organic solvent virus inactivating agent comprises a polyolefin. For example, the above polyolefin fibers, polystyrene particles, and polyacrylate particles. In a preferred embodiment of an embodiment, the polyolefin comprises polystyrene, especially a macroporous adsorption resin comprising polystyrene.

In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises a photosensitizer virus inactivating agent adsorption medium, and the photosensitizer virus inactivating agent adsorption medium contains at least the passivation adsorption medium. Or / and the photosensitizer physically adsorbs the fibers. In the examples of the present invention, the photosensitizer physical adsorbing fibers used include natural fibers, cellulose-based derived fibers, and non-functionalized synthetic fibers.

In an embodiment of the virus inactivation method of the present invention, the photosensitizer physically adsorbing fibers comprise natural fibers. In the examples of the present invention, the natural fibers used include plant fibers, animal fibers (e.g., silk). The plant fibers used include cotton fibers (e.g., non-fat cotton), wood fibers (in filter plates), hemp fibers, pulp (e.g., paper). The fibers also include cellulose based derived fibers (e.g., viscose, acetate, and the like). In an embodiment of the virus inactivation method of the invention, the natural fibers comprise wood fibers.

In an embodiment of the virus inactivation method of the present invention, the photosensitizer physically adsorbing fibers comprise synthetic fibers, preferably non-functionalized synthetic fibers. In an embodiment of the invention, the non-functionalized synthetic fibers used include synthetic fibers other than affinity fibers and ion exchange fibers, such as polyester fibers, polypropylene fibers (eg, ultrafine polypropylene oil absorbing fibers), polyamide fibers, Polyurethane fibers and other polyolefin fibers. In a preferred embodiment, the fiber has one or more of the following characteristics: A). Average diameter 0.01-20 μπι _; Β). Curl degree 1-2, · C). Water absorption 25-50 _g /100 _g fiber. In addition, it is well known that other structural parameters of the fiber, such as the glass transition temperature of the polymer, the electrokinic layer potential of the fiber, and the isoelectric point, etc., also affect its adsorption capacity.

In an embodiment of the virus inactivation method of the present invention, the synthetic fiber comprises a polyolefin fiber. In an embodiment of the invention, certain filter media (e.g., Seitz-SupradurlOO, Seitz-Supradur 200 Seitz-Supradur 500) used as a sensitizing agent for the photosensitizer contain polyolefin fibers.

In an embodiment of the virus inactivation method of the present invention, the virus inactivating agent adsorption medium further comprises particles, and the particles comprise an organic polymer physical adsorbent of a photosensitizer virus inactivating agent. For example, the following treatment system: In one chamber, the photosensitizer virus inactivating agent adsorption medium comprises styrene-containing organic polymer particles and a non-woven fabric or filter of photosensitizer physico-adsorbing fibers (for example, polyolefin fibers) located under the particles. material.

In one embodiment of the virus inactivation method of the invention, the concentration of the photosensitizer virus inactivating agent when the virus is inactivated is from 0.55 to 3.0 μηη _Ο 1 /1. A high photosensitizer virus inactivating agent concentration is beneficial to reduce the time to virus elimination, or / and to apply virus-eliminating conditions that are beneficial to maintain biological activity (eg lower temperature) Degree, lower light intensity, etc.). In fact, the side effect of a solid phase medium containing a viral inactivating agent adsorption medium is one factor limiting the concentration of the photosensitizer virus inactivating agent. In one embodiment, the virus inactivation is carried out at a temperature below room temperature, for example between 4-19.5 ° C, preferably between 15 and 19.5 ° C, more preferably between 15 and 17 ° C. Row.

In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises the organic solvent virus inactivating medium, an organic solvent virus inactivating agent adsorption medium, or/and a photosensitizer physical adsorption fiber. This is a dual virus inactivation method.

In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises an iodine virus inactivating medium, the iodine virus inactivating medium comprising at least iodine, an iodine adsorption medium, and a passivating agent.

In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises a solid phase virus inactivating agent and a passivating agent immobilized thereon. In an embodiment of the present invention, an example of a solid phase virus inactivating agent is a positive charge deep filtration filter (e.g., Cuno Corporation Zetaplus VR filter plate). Among them, the solid phase virus inactivating agent is a solid phase virus inactivating agent based on a positive charge action. Although an example of this embodiment is exemplified in the embodiments of the present invention due to the limitation of the number of existing products, those skilled in the art should know that this method is easy to push on other solid phase virus inactivating agents. .

In an embodiment of the above virus inactivation method of the present invention, the virus inactivating medium has a height greater than 2 cm and less than 6 cm, and the virus inactivating agent adsorption medium has a height greater than 1 cm and less than 3 cm. For safety, it is a well-known practice to use an excess of the adsorption medium, however, excessive adsorption medium increases the risk of side effects.

In an embodiment of the virus inactivation method of the present invention, the solid phase medium comprises a depth filter medium, the deep filter medium containing the virus inactivating agent adsorption medium, and having the following characteristics: A) The average density is greater than 0.25 g/cm ³ ; and B). The ash content is less than 1%. According to an embodiment of the invention, controlling ash has important implications for minimizing side effects. The filter medium may contain, in addition to the fibers, an adhesive, a reinforcing agent (e.g., polyolefin), and the like. Higher density filter media can have higher adsorption capacity. In one embodiment, the solid phase medium comprises one of the following filters or their analogs: Seitz-BiolO, Seitz-Bio20, Seitz-bio40, Seitz-SupradurlOO, Seitz-Supradur200, Seitz-Supradur500, Seitz -Supradurl000. In a preferred embodiment, the average density of the filter media is greater than

Seitz-SupradurlO0, Seitz-Supradur 200) _o In an embodiment of the method of the invention, the depth of the depth filter media is greater than 6 mm.

In the present invention, the passivating adsorption medium may contain one or more passivating agents, one or more adsorbing media, and the like. The adsorption medium in the passivation adsorption medium includes: a virus inactivating agent adsorption medium, a virus inactivating agent adsorption medium in the virus inactivating medium, and a leukocyte removal medium. In an embodiment of the present invention, the virus inactivating agent adsorption medium contains one or more of the following adsorbents: activated carbon, inorganic adsorbent, Organic polymer chemical adsorbent, organic polymer affinity adsorbent, organic germanium molecular physical adsorbent. The organic polymer affinity adsorbent includes particles or fibers to which a ligand (for example, a C6-C18 carbon chain, a hydrazine-containing end group or a viral RNA, or a DNA replica) is immobilized. These adsorbents have an adsorption function for virus inactivating agents, but tend to have strong undesired reactivity.

In an embodiment of the virus inactivation method of the present invention, the adsorbing medium in the passivating adsorption medium comprises activated carbon. In the embodiments of the present invention, activated carbon occurs in various forms such as activated carbon powder, activated carbon felt, activated carbon-containing filter material, and the like.

In an embodiment of the virus inactivation method of the present invention, the adsorbing medium in the passivating adsorption medium comprises an ion exchange adsorption medium. The ion exchange adsorption medium includes: an inorganic adsorbent such as diatomaceous earth, perlite, glass fiber, etc.; an organic polymer chemical adsorbent such as an ion exchange resin, an ion exchange fiber, or the like.

In an embodiment of the virus inactivation method of the present invention, the adsorption medium in the passivation adsorption medium comprises the above physical adsorption medium (organic solvent virus inactivating agent adsorption medium, photosensitizer physical adsorption fiber). Passivation of the physical adsorption medium makes it possible to further reduce side effects.

In the embodiment of the present invention, the passivating agent is preferably an organic substance that can be injected into a human body. These passivating agents are characterized by: 1) themselves being relatively inert; 2) can be immobilized on a solid phase medium to inhibit its side effects without meaningful adverse effects on its function; 3). The controlled amount of passivating agent is detached from the solid phase medium or other composition into the biological fluid, which does not cause substantial damage to the safety of use of the biological fluid.

In an embodiment of the virus inactivation method of the present invention, the passivating agent in the passivation adsorption medium comprises a hydrophilic group or/and a lipophilic group, and the solid phase medium pair can be lowered An organic substance that describes the side effects of biological fluids. An organic substance having an oleophilic group is poorly water-soluble (for example, a natural oil, an organic solvent virus inactivating agent), and a considerable number of organic substances having a hydrophilic group or having a hydrophilic group and a lipophilic group are considered to have poor adsorption force ( For example, surfactants, hydroxy compounds, amino acids), no passivating agents are used as solid phase media. Surprisingly, such materials are useful as passivating agents in the practice of the present invention and are intended to serve the objectives of the present invention.

In an embodiment of the invention, the passivating agent content in the solid phase medium is greater than

It is preferably greater than 1 μιηο1/αη ³ . For the treatment system of the present invention, the purpose of the present invention (e.g., minimizing side effects) can be met only in certain cases when the level of passivating agent is suitably large. Some unexpected results in the examples of the present invention are that the adsorption of viral inactivating agents (e.g., methylene blue) is still sufficiently strong when the amount of certain passivating agents on the activated carbon material is greater than 5 μηιο1/οιη ³ . As for the upper limit of the content of the passivating agent, it should be different according to its activity (specific adsorption capacity or virus inactivation ability) and the degree of passivation required (for example, the degree of passivation of biological fluids with or without clotting factors may be different) , optimized according to known methods.

In an embodiment of the virus inactivating method of the present invention, the purifying agent comprises a natural fat. Place Natural oils and fats include natural oils and natural oils and fats. These materials all have an oleophilic group. In the embodiment of the present invention, the natural fats and oils include vegetable oils and phospholipids, wherein: vegetable oils include castor oil, soybean oil, tea oil, and glycerin; and phospholipids include cephalin, lecithin, and plant phospholipids. Examples of natural oil and fat derivatives include vegetable oil emulsions, fat emulsions, and the like.

In an embodiment of the virus inactivation method of the present invention, the purification agent comprises a hydroxy compound. The hydroxy compound has a hydrophilic group including alcohols (for example, ethanol, glycerin, glycerin sodium chloride, and the like), saccharides, or/and their respective derivatives. In one embodiment, the saccharide comprises a monosaccharide, a polysaccharide, or/and a polypolysaccharide, for example: glucose, mannitol (Molecular Formula C ₆ H ₁₄ 0 ₆ ), Sorbitol, Glycerol Fructose, Sucrose, Hydroxyethyl Starch, lentinan, etc.

In an embodiment of the virus inactivation method of the invention, the purification agent comprises an amino acid. The amino acid has a hydrophilic group including cystine, lysine, tyrosine, glycine, arginine, valine, serine, a polymer of these amino acids, and the like. The surprising result of embodiments of the present invention is the saccharide or /amino acid and can be adsorbed on the surface of the activated carbon and reduce the side effect of the activated carbon on the biological fluid. Polyamino acids also have a similar effect.

In an embodiment of the virus inactivation method of the present invention, the purification agent comprises an organic solvent. In the embodiment of the present invention, the organic solvent includes an organic solvent virus inactivating agent such as tributyl phosphate (TnBP), diethyl ether, glycerin, ethyl carbonate, ethyl lactate, benzyl benzoate, and the like.

In an embodiment of the invention, the passivating agent comprises a polypeptide. The polypeptide has hydrophilic groups such as albumin, serum, and milk, and their respective derivatives.

In an embodiment of the invention, the passivating agent comprises a surfactant. The surfactant has a hydrophilic group and a lipophilic group. The present inventors have found that a large number of surfactants, especially surfactants having both hydrophilic groups and lipophilic groups in the molecular structure, can be adsorbed on the surface of activated carbon and reduce the side effects of activated carbon on biological fluids. Hydrophilic groups include: -OH, -OS0 ₃ , - (C3⁄4CH ₂ 0) ₃ , -N(CH ₃ ) ₂ , -N(CH ₃ ) ₃ , -CH ₂ COO, -NH _{2 ;} lipophilic groups Including: organic ring groups, -(CH ₂ i-, etc. Examples of surfactants include: Triton-based surface active substances, Tween-based surface active substances, sodium cholate, tetrahydrofurfural polyglycol ether, two Methylacetamide, polyvinylpyrrolidone, and dimethyl sulfoxide. Examples of partial surfactants as passivating agents are given in the examples.

In an embodiment of the virus inactivation method of the present invention, the virus inactivating medium is immobilized with an organic solvent virus inactivating agent and an organic solvent deactivator, and the organic solvent deactivator and the organic solvent virus are inactivated. The total content of the agent is greater than 0.2 μπιο1/∞ι ³ , preferably greater than 0.3 μιηο1/αη ³ . The examples of the present invention show that only when the content of the organic solvent virus inactivating agent (for example, ΤηΒΡ) is greater than a certain degree, the organic solvent can be used as a virus inactivating agent for effective virus inactivation and as a passivating agent. Minimize side effects. The embodiment of the present invention demonstrates that the content of the organic solvent virus inactivating agent is greater than 0.2 mol/cm ³ . In The solid phase medium may also contain an appropriate amount of detergent D (e.g., greater than 0.01 mol/cm ³ ) or other passivating agent (e.g., polysaccharide). When the D content is sufficiently large (for example, greater than 0.2 Mmol/cm ³ ), D can also be used as both a virus inactivating agent and a passivating agent.

In an embodiment of the invention, the passivating agent comprises a combination of two or more of the above passivating agents, for example: dextran 40 / glucose, dextran 40 / albumin, albumin / glucose, TnBP / spit Wen-80/glucan, and so on.

In one embodiment of the virus inactivation method of the invention, the treatment system further comprises a leukocyte-free solid phase medium. For example, an embodiment of the method of the present invention comprises at least: A) providing a single blood component; B) providing said treatment system, said treatment system further comprising a leukocyte solid phase medium; C). Flowing the biological fluid through the treatment system for leukocyte removal and virus inactivation or/and viral inactivating agent adsorption; D). The treatment system from C) is substantially free of the virus inactivating agent and A single blood component of white blood cells. In an embodiment of the method of the present invention, the leukocyte-removing solid phase medium comprises the organic polymeric physical adsorbent.

In an embodiment of the virus inactivation method of the present invention, the leukocyte-removing medium medium comprises the virus inactivating agent adsorption medium.

In one embodiment of the virus inactivation method of the invention, the biological fluid comprises a single human blood component. In an embodiment of the virus inactivation method of the present invention, the content of the organic solvent virus inactivating agent in the organic solvent virus inactivating medium is equal to or less than 3 X (10 ppm of the volume of the single blood component) Ml), or less than (lOppmX the volume of the single-part blood component ml). In fact, limiting the amount of organic solvent virus inactivating agent may help to eliminate the detection of organic solvent virus inactivating agents in the final product.

A treatment system for effectively reducing virus hazards of the present invention is the treatment system described in the above virus inactivation method.

An apparatus for effectively reducing virus hazards of the present invention, comprising at least the above-described processing system. The device includes a column, a filter, a kit, and the like. In an embodiment of the invention, some of the devices further comprise a viral inactivating agent addition structure, or/and a virus inactivation reaction site, or/and a wash solution.

In one embodiment of the device of the present invention, the device is a single human blood component treatment kit. Example

The invention will be described in more detail below with reference to examples. However, it should be understood that the embodiments of the invention are merely illustrative of the individual aspects of the embodiments of the invention. Those skilled in the art will appreciate that the present invention is not limited to these given modes of implementation (e.g., given processes, parameters, and combinations). Because the content of the invention is clear, the specific processes, parameters and combinations can be varied. In the following examples, the reagents and materials used were all commercially available reagents and materials. Among them, the adsorption media used are:

1) Activated Carbon: The activated media for the activated carbon-containing virus inactivating agent are activated carbon powder, activated carbon felt (ZC-1200A, China Zichuan Carbon Fiber Co., Ltd.) and activated carbon filter plates (AKS 5 and AS 6, respectively). Seitz, Germany).

2). Ion exchange adsorbent: The virus inactivating agent adsorption medium containing the ion exchange adsorbent is silicon oxide particles and glass fibers, respectively. The material containing the silicon oxide particles was a perlite-containing filter plate (Zetaplus Delipid, Cuno Corporation).

3 Affinity adsorbent: The virus inactivating agent adsorption medium containing the affinity adsorbent is C18 reverse phase gel (Water Company, USA).

4). Physical adsorption medium. Among them, the physical adsorption media used are:

(1) Photosensitive agents Physically adsorbed fibers, including natural fibers, cellulose-based derived fibers, and non-functionalized synthetic fibers. The natural fibers used include plant fibers, animal fibers (e.g., silk). The plant fibers used include cotton fibers (e.g., non-fat cotton), wood fibers (in filter plates such as Seitz Bio series filter plates), and pulp (e.g., paper). The non-functionalized synthetic fibers used include synthetic fibers other than affinity fibers and ion exchange fibers, such as polyester fibers, polypropylene fibers (for example, ultrafine polypropylene oil absorbing fibers), polyamide fibers, and polyurethane fibers (both non-woven fabrics). ). In particular, polyolefin fibers are contained in the filter material used as the sensitizer physical adsorption medium in the following examples, such as Seitz Supradur 80, Seitz Supradur 500 Seitz Eco 1000, and the like.

(2) Organic solvent physical adsorption medium, including fibers (organic solvent physical adsorption fibers) and macroporous adsorption resins (organic solvent physical adsorption resins). The fiber used contains an organic polymer physical adsorbent containing an organic solvent virus inactivating agent, such as natural fibers (for example, Seitz Bio series filter plates), polyolefin fibers (such as Seitz Supradur 80, Seitz Supradur 500, Seitz Eco 1000, etc.), and many more. The macroporous adsorption resin (organic solvent physical adsorption resin) used includes polystyrene particles (for example, Amberlite XAD-7HP, Amberlite XAD-16), phenolic polycondensation type particles, polyacrylate particles, and polyalkyl ester-containing macroporous adsorption resin. Amberlite XAD- 7HP is a polar polystyrene macroporous resin particle, and Amberlite XAD-16 is a non-polar polystyrene macroporous resin particle. The particles used have one or more of the following characteristics: A). Average particle size 5-1000 μιη _; Β). Specific surface area 100-2000 m dry powder; C). Average pore diameter less than 5-600 A; D) Excluding volume less than 50,000 molecular weight . The porous protein solid phase medium used in part has a spherical protein exclusion lower limit molecular weight of less than 5,000.

In the following examples, the photosensitizer physical adsorbing fiber or the organic solvent physically adsorbing fiber comprises a fiber filament, a nonwoven fabric, a cloth, and a depth filter medium. The deep filter media containing the photosensitizer physically adsorbing fibers or organic solvent physically adsorbing fibers, the average density is greater than 0.25 g/cm ³ , and the ash content is less than 1%, including various German Seitz products (KS80, K900, Supradur 80 > P30) , Eco 1000, Permadur 0/400A, T2600, Bio20, Bio40, Bio60> Supradur 100, etc.). In the present invention, the prefix Seitz- refers to the product of the German company Sdtz. Professionals should be aware that similar products from other companies have similar adsorption properties. The permeability of some of the deep filter media used in the solid phase media is less than 200 L/m ² min, which are: Seitz-Supradur 100, Seitz-biolO, Seitz-bio 40 ₀

In the following examples, the passivating agent used includes:

1) A natural fat or oil having an oleophilic group or/and an organic solvent, for example, castor oil, soybean oil, tea oil, fat milk, tributyl phosphate (abbreviated as ΤπΒΡ in the present invention), and diethyl ether.

2) Surfactants having a hydrophilic group and an oleophilic group, for example: sodium cholate, Triton® 100, Tween 80, and polyvinylpyrrolidone.

3) Hydrocarbyl compounds having a hydrophilic group, such as: glycerin, glucose, maltose, dextran (dextran), water-soluble cellulose, mannitol (C ₆ H ₁₄ 0 ₆ ), sorbitol, hydroxyethyl Starch, lentinan.

4) . Amino acids having a hydrophilic group, such as: valine, arginine and glycine.

5). Polypeptides with hydrophilic groups, such as: human albumin (Beijing Tiantan Biological Products Co., Ltd.), Buxuekang (Germany Biotest) and milk.

6). A composite passivating agent for the above passivating agent, for example: dextran 40/glucose, dextran 40/albumin, albumin/glucose, TnBP/Tween-80/dextran. In accordance with the following examples of the invention, those skilled in the art will readily be able to use other similar passivating agents for the purpose of passivation.

In the following examples, the virus inactivating agent used includes:

1). Organic solvent virus inactivating agent or organic solvent virus inactivating agent/detergent, for example. · Tributyl phosphate, diethyl ether, TnBP/Tween-80, TnBP/Triton X100, TnBP/sodium cholate, diethyl ether /Tween 80.

2) Photosensitizer virus inactivating agents, such as dye virus inactivating agents or psoralen virus inactivating agents. The dye-like virus inactivating agents used were methylene blue, methyl violet, and toluidine blue, respectively. The psoralen virus inactivating agents used were psoralen and 8-m _e th _OX ypr _OT al _{en , respectively} . It should be understood by the skilled person that other psoralens have almost the same adsorption properties as the psoralen used.

3) . Iodine. Especially iodine-PVPP filter plates (Sdtz).

4) . Solid phase virus inactivating agents, such as Zetaplus VR filter (Cuno:).

In the following examples, the conditions for virus inactivation are generally as follows: pH is 2.0-12.0; temperature is -5 to 6 CTC; amount of solid phase medium and biological liquid containing virus inactivating agent according to virus inactivation experiment Preferably (for example, solid medium volume / biological liquid volume between 1% and 30%); virus inactivation hydrodynamic conditions are preferred according to virus inactivation experiments (eg linear flow rate 0.1-lOcm/min, pressure 0.1-) 5 kg/cm ² ); the amount of solid phase medium and biological liquid containing the virus inactivating agent adsorption medium is preferably according to the virus inactivating agent removal experiment (for example, the solid medium volume/bioliquid volume is between 1% and 30%) Between) The determination of the liquid phase adsorption reaction conditions is preferably carried out in accordance with a well-known rule, for example: the greater the flow rate of the biological liquid, the smaller the adsorption efficiency;

In the following examples, the virus inactivation efficiency was carried out according to a known method using a pattern lipid enveloped virus.

VSV (initial titer greater than 10 ⁴ · ⁸ ) and Sendai virus (initial titer greater than 10 ⁵ · ¹ ) were studied. In the following examples, the detection of various biological activities was carried out by a known method. In the present invention, the leukocyte removal rate is measured and calculated by a known method.

The devices prepared in the following examples include devices that can be used to effectively reduce the pathogen hazard of a single human blood component. It comprises at least the treatment system of the invention, wherein the chamber is a conventional filter cartridge or chromatography column. The filter prepared by the following examples effective in reducing the pathogen hazard of a single human blood component, generally, the inner diameter is

3cm, the solid phase media loading height is 3-5cm, and there is a 0.4μηι filter at the outlet end of the filter to prevent the media debris from leaking out. The column prepared by the following examples is effective for reducing the pathogen hazard of a single blood component, generally having an inner diameter of 1 cm, a solid phase medium loading height of 3-5 cm, and a 0.4 μm filter at the outlet end of the column. In case the media debris leaks out.

Example 1 Preparation of passivated solid phase medium of the present invention

The following examples give a few more detailed examples.

Example 1.1 Virus inactivating agent adsorption medium containing passivating agent

The solid phase medium prepared in this example is an adsorption medium/passivator composite.

1. 1.1 Preparation method

i). Preparation of passivation agent dispersion treatment system

In this embodiment, the passivating agent is at a preferred concentration (for example, natural fat: 0.2-0.5%; organic solvent: 0.3-2.0%; surfactant: 1.0-3.0%; polypeptide: 1.0-10.0%; amino acid: 3.0 -10.0%; etc.) Disperse in an aqueous solution (eg PBS buffer). However, it is to be noted that other media (e.g., organic solvent virus inactivating agents) may also be used as a passivating agent dispersion medium to prepare a passivator dispersion treatment system (e.g., solution, suspension) depending on the nature of the passivating agent used. At this time, the concentration of the dispersion treatment system (w/v) may be from 0.1% to 50°/. between.

The dispersion of the low water-soluble organic substance deactivator can be carried out by a known technique. There are many methods for dispersing natural fats and oils in aqueous solutions, such as emulsification, air removal, and the like. In one embodiment of this embodiment, a surfactant is also added to the dispersion medium. The use of surfactants (eg Tween 80, Triton X 100) in addition to facilitating the dispersion of the above-mentioned passivating agents, in particular natural oils or/and organic solvent virus inactivating agents, in water, sometimes as an eluent The adsorption of the above passivating agent on the adsorption medium is controlled.

(2) Fixing the passivating agent to the adsorption medium

In this embodiment, the passivating agent is fixed on the adsorbent medium, or substantially by adsorption. However, it is important to know that other fixing methods (such as covalent bonding) can also be selected. For example, the liquid passivating agent can also be directly contacted with the adsorption medium to carry out the adsorption reaction. If a powdery adsorption medium (for example, activated carbon) is used, it is added to the above-prepared passivator dispersion system for adsorption reaction. If a bulk adsorption medium (for example, an activated carbon filter plate) is used, the passivation agent dispersion system prepared above is a mobile phase, and the adsorption medium is used as a stationary phase for a flow adsorption reaction.

All binding reactions were carried out using optimized passivator/adsorption media ratios under optimized reaction conditions. These optimizations are performed in accordance with well-known bonding techniques (e.g., adsorption techniques). Conditions for the adsorption reaction include: reactant addition amount, pH, temperature, time, concentration of certain additives (e.g., surfactant, salt, etc.), mobile phase flow rate (at the time of flow adsorption reaction), and the like. Those skilled in the art will recognize that by controlling these conditions, the adsorption reaction is controlled to achieve the desired result (e.g., amount of adsorption). The uniformity of the adsorption reaction is also a priority.

Those skilled in the art will recognize that the adsorbent/passivator complex can also be prepared by immobilizing a viral inactivating adsorbent (e.g., C18) on a solid support (e.g., chromatography gel) followed by immobilization of the passivating agent.

(3). Cleaning unconjugated

Passivation agents and other substances that are not fixed or weakly adsorbed on the adsorption medium, and may be washed or prevented by different washing liquids (for example, PBS buffer, preferred concentration of urea solution, alcohol solution, etc.) according to different needs. Sexual washing.

Some of the solid phase media prepared in this example are listed in Table 1.

The composite prepared above, especially the particulate matter, can be used alone as a solid phase medium (for example, A5-A30 in Table 1), or with other components (such as a speed increasing substance, a binder, a solubilizing agent, etc.). After mixing, it is used as a solid phase medium containing an adsorption medium and a passivating agent. For example, in Table 1, A1 adsorption medium/passivator complex and 10% by volume chromatographic gel (Sepharose FF, product of Pharmacia) are mixed; A2 contains adsorption medium/passivator complex and 10% by volume Perlite mix. Other components in the solid phase medium (eg, speed-increasing substances, binders, solubilizers, etc.) can be passivated according to actual needs according to the undesired reactivity of the medium (for example, forming other solid phases) The component/passivator complex), or after mixing with the adsorption medium, is passivated (eg, forming a solid phase media/passivator complex:). Their passivation method is the same as that in the above-described method for preparing an adsorbent medium/passivator compound. In Table 1, A1 is prepared by first forming a solid phase component/passivator complex and then mixing to form a solid phase medium; A2 is first mixed to form a solid phase medium, and then a passivating agent is added to passivate the different components. Prepared.

Table 1. Part of virus inactivating agent adsorption medium containing passivating agent

Composite adsorption medium passivator composite adsorption medium passivation agent

A1 activated carbon powder castor oil A21 AKS5 Tween -80

A2 activated carbon powder dextran A22 AKS5 polyvinylpyrrolidone

A3 ZC-1200A Human Albumin A23 AKS5 TnBP

A4 AKS 6 Proline A24 AKS5 Castor Oil A5 AKS 6 arginine A25 A S5 soybean oil

A6 AKS 6 glycine A26 AKS5 fat emulsion

A7 AKS 6 human albumin A27 AKS5 TnBP/ Triton XI 00

A8 AKS 6 dextran A28 AKS5 dextran 40/glucose

A9 AKS 5 human albumin A29 A S5 dextran 40/albumin

A10 AKS 5 Biankang A30 AKS5 TnBP/Tween-80/Glucan

All AKS 5 arginine A31 Amberlite dextran 40/glycine

XAD-

7HP

A12 AKS 5 glycine A32 Amberlite dextran 40/glycine

XAD-1

A13 AKS 5 Glycerin A33 Glass fiber Human albumin

A14 AKS 5 Glucose A34 Polyester Fiber Human Albumin

A15 AKS5 Dextran A35 Polyurethane Fiber Human Albumin

Wei

A16 AKS5 malt, sugar A36 Zetaplus human albumin

Delipid

A17 AKS5 Mannitol A37 Zetaplus Dextran 40/Albumin

Delipid

A18 AKS5 Lentinus Polysaccharide A38 C18 Gel Human Albumin

A19 AKS5 polyethylene glycol A39 C18 gel dextran 40/albumin

A20 AKS5 Triton A40 PVPP Filtration Human Albumin

X100 board

In Table 1, some of the virus inactivating agent adsorption media containing the passivating agent are both an organic solvent virus inactivating agent adsorption medium and a photosensitizer virus inactivating agent adsorption medium (for example, those containing activated carbon and macroporous adsorption resins). .

1.1.2 Identification

(1) . Determination of the content of passivating agent in solid phase media

In this embodiment, the passivating agent content of the solid phase medium = (total amount of passivating agent added - unfixed passivating dose) / amount of solid phase medium. Among them, the unfixed passivation dose is obtained by well-known correlation measurement techniques. For example, the method for measuring natural fats and oils is a spectrometer method; the method for measuring an organic solvent virus inactivating agent is gas chromatography; the method for measuring a surfactant is gas chromatography; and the method for determining a polypeptide is high pressure liquid chromatography; The method of determination is high pressure liquid chromatography; and the like. In the solid phase medium prepared in this example, in most cases, the content of the passivating agent is greater than 0.05 μπιο1/αη ³ and individually greater than 0.4 mmol/cm ³ .

(2) . Determination of specific adsorption of solid phase media

In the present invention, the specific adsorption amount of the virus inactivating agent = (the total amount of the virus inactivating agent added - the unadsorbed virus inactivating dose y the volume of the solid phase medium).

In this embodiment, the organic solvent virus inactivating agent (TnBP) and the photosensitizer virus inactivating agent (methylene blue) are divided into Do not be used to determine specific adsorption. The measurement of methylene blue uses a well-known spectrophotometer method. The measurement of TnBP uses a well-known gas chromatography. The amount of adsorption of the virus inactivating agent is measured by a known method for measuring the dynamic adsorption amount. The apparatus used for the measurement was a column container (volume 10 ml) or a filter (volume 10 ml) respectively containing the solid phase medium and the control medium prepared in the present example, and 100 ml of the virus inactivating agent solution (methylene blue concentration 100 g/ml) was measured. The TnBP concentration of 10 mg / ml was passed through the apparatus at a linear velocity of 0.3 cm / min.

In Table 1: The adsorption amount of the solid phase medium A1-A35 to the photosensitizer virus inactivating agent (methylene blue) is greater than O.Olmmol/cm ³ , and the individual is greater than 0.02 mmol/cm ³ , and sometimes even greater than 0.1 mmol/cm. ³ (for example, Al, A2, A32, etc.); the adsorption amount of the organic solvent virus inactivating agent (TnBP) of the solid phase media A1-A30 and A36-A39 is greater than 0.05mmol/cm ³ , and the individual is greater than O.lmmol/cm ³ , individual even greater than 0.3mmol / cm ³ ; A1-A30 adsorption of iodine greater than lmmol / cm ³ . In addition, some of the above solid phase media (especially those containing activated carbon) have similar results for the specific adsorption of other photosensitizer virus inactivating agents, such as psoralen.

In Table 1, A33-A35 can also be used as a leukocyte-removing medium.

(3) Determination of side effects of solid phase media

In this example, the measurement of side effects includes measuring the amount of non-specific adsorption or / and APTT (human plasma fraction prothrombin activity). In this example, the amount of non-specific adsorption = (indicating the total amount of reagent added - the total amount of indicator reagent not adsorbed) / the volume of the solid phase medium.

In this example, human albumin (sample C) (Tiantan Biological Products Co., Ltd.) was used as a non-specific adsorption indicating reagent; partial prothrombin activity time of human plasma (sample D) (abbreviated as APTT in the present invention) Used as an indicator of changes in the coagulation system. The sputum kit was purchased from the Chengdu Institute of Blood Transfusion, Chinese Academy of Medical Sciences.

As in the above-described measurement method of specific adsorption, the measurement of side effects is also carried out by a known dynamic reaction measurement method. The ratio of human albumin (5%) or human plasma (5.5% protein) to the solid medium was between 3:1 and 5:1.

Compared with the control adsorption medium, the adsorption medium/passivator composite prepared in this example has a significant decrease in side effects, which is reflected as: A). The albumin adsorption amount (mg/cm ³ ) decreases by more than 25%, and the individual decreases by more than 50%. (: for example, albumin/adsorption medium complex, vegetable oil/adsorption medium complex, sugar/adsorption medium complex, etc.); B). The increase in human plasma APTT is more than 30%, and the individual drop is more than 100% ( For example, albumin/adsorption medium complex, vegetable oil/adsorption medium complex, sugar/adsorption medium complex, etc.).

Example 1.2 Virus inactivating medium containing passivating agent (1)

The solid phase medium prepared in this example is an organic solvent virus inactivating agent/adsorption medium/passivator compound.

1.2.1 Preparation method (1) Preparation of virus inactivating agent/adsorption medium complex from adsorption medium

(A). Preparation of a virus inactivating agent solution or suspension

In this embodiment, a virus inactivating agent solution or suspension is prepared according to a known technique using PBS buffer as a dispersed phase, for example: TnBP/Triton X 100/water solution (5% TnBP, 5% Triton X 100); TnBP/spray Warm 80/water solution (3% TnBP, 10% Tween 80 concentration); β-propiolactone/Tween 80/water solution (0.5% β-propiolactone);

(Β). Fix the virus inactivating agent to the adsorption medium

The virus inactivating agent in the liquid phase is adsorbed on the solid phase adsorption medium by a conventional solid-liquid phase adsorption reaction condition.

(C). Fixing the passivating agent to the adsorption medium

The fixing method is the same as the method of "fixing the passivating agent to the adsorption medium" in the embodiment 1.1.

(2) . Preparation of virus inactivating agent / adsorption medium / passivation complex from virus inactivating agent / adsorption medium complex

The preparation method of the virus inactivating agent/adsorption medium complex in this embodiment is the same as the method of "fixing the virus inactivating agent to the adsorption medium" in the present embodiment (1) (e.g., iodine-PVPP filter plate). The method of fixing the passivating agent to the virus inactivating agent/adsorption medium composite is the same as the method of "fixing the passivating agent to the adsorption medium" in the embodiment 1.1.

Some of the solid phase media prepared in this example are listed in Table 2. The organic solvent virus inactivating agent/adsorption medium/passivator complex (for example, B1-B3 in Table 2) in this embodiment may be used alone or in combination with other components (for example, a speed increasing substance, a binder, The solubilizer, etc.) is used as a solid phase medium containing the virus inactivating agent after mixing. The passivation method may also be first mixed to form a solid phase medium, followed by passivation to passivate, or passivate the different components and then mix them.

1.2.2 Identification

Virus inactivation medium prepared in the present embodiment, the measured efficiency of inactivated virus into which ^ ¹ described above, measurement was carried out at room temperature, less than the linear flow rate of lml / cm / min, the volume of biological fluid with a solid support is less than 100. The virus inactivating agent/adsorption medium/passivator complex prepared in this example has an effective virus inactivating function.

The virus inactivating medium prepared in this example has the same method of determining the passivating agent content, the virus inactivating agent content, and the solid phase medium side effect as in the corresponding method in the first embodiment.

The virus inactivating medium prepared in this example in Table 2 has a passivating agent content of more than 0.05 mol/cm ³ and an individual content of more than 0.4 mmol / cm ³ . Compared with the control virus inactivating agent/adsorption medium complex, the virus inactivation efficiency was not significantly changed by passivation, but the side effects were significantly decreased: A). The albumin adsorption amount (mg/cm ³ ) decreased by more than 15%, and decreased individually. More than 30%; B). The increase in human plasma APTT is more than 20%, and the individual is reduced by 50%. the above.

In particular, in B7 or B8 of Table 2: A). The virus inactivating agent comprises an organic solvent virus inactivating agent; B). the passivating agent comprises an organic solvent virus inactivating agent; and C The content of the organic solvent virus inactivating agent in the solid phase medium is greater than 0.1 mmol / cm ³ , or even greater than 0.3 mmol / cm ³ (for example, 0.40 mmol / cm ³ ) _{0 at} this time, the organic solvent virus inactivating agent can It can be used as a virus inactivating agent for effective virus inactivation, and can also be used as a passivating agent for a virus inactivating agent adsorption medium (for example, activated carbon) to minimize side effects. Example 1.3 Virus inactivating medium containing passivating agent (2)

In this example, a solid phase virus inactivating agent/passivator complex was prepared. The solid phase virus inactivating agents used were Cuno-Zetaplus VR virus-killing filter plates and Seitz-iodine-PVPP filter plates. The method of fixing the passivating agent to the solid phase virus inactivating agent is the same as the method of "fixing the passivating agent to the adsorption medium" in Example U. The preparations are listed in Table 2.

The virus inactivating medium prepared in this example has the same method for determining the virus inactivation efficiency, the depressant content, and the side effects, respectively, in accordance with the corresponding measurement methods of the examples.

The solid phase virus inactivating agent/passivator complex prepared in this example in Table 2 has a passivating agent content of more than 0.05 μπιο1/ιι ³ . Compared with the control solid phase virus inactivating agent, the virus inactivation efficiency was not significantly changed by passivation, but the side effects were significantly decreased: Α). The albumin adsorption amount (mg/cm ³ ) decreased by more than 5%; B). Human plasma The APTT rise is down more than 10%.

Table 2. Part of solid phase media containing virus inactivating agent

Complex adsorption medium virus inactivating agent passivator

B1 activated carbon powder S/D arginine

B2 activated carbon powder S/D proline

B3 activated carbon powder S/D glycine

B4 AKS5 S/D Human Albumin

B5 A S5 S/D milk

B6 AKS5 S/D Bianxikang

B7 AKS5 S/D TnBP

B8 AKS5 S/D TnBP/ Triton XI 00

B9 AKS5 S/D Triton XI 00

B10 AKS5 S/D Glucose

B11 AKS5 S/D dextran

B12 PS adsorption resin S D dextran 40/glucose

B13* PVPP iodine human albumin

B14 PS** adsorption resin S/D dextran 40/glucose

B15 Zetaplus VR filter human albumin B16 Zetaplus VR filter material glycine

B13* _: SEITS PVPP iodine filter plate; PS**: polystyrene

Example 1. 4 virus inactivating agent adsorption medium containing deactivating agent / leukocyte solid phase medium

In this embodiment, the leukocyte-removing medium-virus inactivating agent adsorption medium used is: glass fiber, cotton fiber, polyester fiber, polyurethane nonwoven fabric, Seitz-Supradur 100 polypropylene fiber filter plate The passivating agent used is the above passivating agent (for example, glycine, dextran 40/glucose, and mixtures thereof).

The passivating agent/viral inactivating agent adsorption medium-de-whitening cell solid phase medium composite prepared in this embodiment is prepared in the same manner as the adsorption medium/passivator complex in the embodiment 1.1, and the adsorption reaction is in the leukocyte-removing cell. The medium-viral inactivating agent is adsorbed between the adsorption medium and the passivating agent, and the method for determining the content of the passivating agent and the side effect of the solid phase medium is the same as that of the corresponding method in Example 1.1, respectively.

The preparation of the present embodiment has a leukocyte removal rate of over 99%, and a methylene blue removal rate of over 95%, and the side effects thereof are significantly lower than the corresponding leukocyte-removing medium without a passivating agent: A). Albumin The amount of adsorption (mg/cm ³ ) decreased by more than 15%; B). The increase in human plasma APTT decreased by more than 20%.

Example 2. Preparation of the apparatus of the present invention (1)

The apparatus prepared in this example, the solid phase medium used is selected from the passivated solid phase medium prepared in Example 1, especially the examples 1.1-1.4 therein.

The apparatus of this embodiment is obtained by two methods: A) loading a prepared passivated solid phase medium into the chamber; B) loading the solid phase medium into the chamber and then passivating it with a passivating agent, The passivation method is the same as the method of "fixing the passivating agent to the adsorption medium" in Example 1.

The apparatus prepared in this embodiment has the same identification method as the corresponding identification method of the passivated solid phase medium in Embodiment 1. In general, its functions (e.g., specific adsorption capacity, leukocyte removal rate, virus inactivation efficiency) and side effects are consistent with the function and side effects of the passivated solid phase medium contained therein, respectively. That is, the function is not significantly reduced by the addition of a passivating agent, and its side effects are significantly reduced.

The following examples give a few more detailed examples.

Example 2.1 Virus inactivating agent removal device

Example 2.1.1 Virus inactivating agent removal filter

In this embodiment, the solid phase medium in the filter is selected from the group consisting of a passivating agent/viral inactivating agent adsorption medium (fiber or/and filter medium). For example, passivating agent/polystyrene fiber composite, passivating agent/polypropylene fiber composite, passivating agent/activated carbon filter plate composite, and the like.

Example 2.1.2 Virus inactivating agent removal column

In this embodiment, the solid phase medium in the virus inactivating agent removal column is selected from the group consisting of a passivating agent/viral inactivating agent. Medium (particle) composite. For example, passivating agent/polystyrene macroporous adsorption resin composite, passivating agent/activated carbon powder composite, and the like.

Example 2.2 Virus inactivation device

Example 2.2.1 Virus inactivation filter

In this embodiment, the solid phase medium in the virus inactivating filter is: 1) a virus inactivating medium immobilized with a passivating agent (for example, a high content of TnBP/earth temperature 80/activated carbon filter plate composite); a virus inactivating medium immobilized with a passivating agent and a virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, a high content of TnBP/earth temperature 80/activated carbon filter plate complex and a sugar-amino acid purifying agent/activated carbon) a filter plate complex; and 3) a virus inactivating medium and a virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, TnBP/earth temperature 80/activated carbon filter plate complex and sugar-amino acid purifying agent/activity Carbon filter plate composite :).

Example 2.2.2 Virus inactivation column

In this embodiment, the solid phase medium in the virus inactivation column is: 1) a virus inactivating medium immobilized with a passivating agent (for example, a high content of TnBP/earth temperature 80/styrene-containing polymer adsorption resin composite) 2). Virus inactivation medium with passivator immobilized and virus inactivating agent adsorption medium with or without passivating agent (eg high content TnBP/earth temperature 80/styrene-containing polymer adsorption resin complex) And sugar-amino acid purifier/activated carbon filter plate composite, or high content TnBP/earth temperature 80/styrene-containing polymer adsorption resin composite and styrene-containing polymer adsorption resin :); and 3). virus Inactivation medium and virus inactivating agent adsorption medium to which a passivating agent is immobilized (for example, styrene-containing polymer adsorption resin/TnBP/earth temperature 80 composite and passivating agent/styrene-containing polymer adsorption resin composite) .

Example 2.3. Leukocyte-removing/viral inactivating agent removal filter

In this embodiment, the solid phase medium in the leukocyte-removing filter is selected from the group consisting of a passivating agent / a leukocyte-removing medium / a virus inactivating agent adsorption medium complex. For example, a filter cartridge is packed with a T 2600 filter plate, 4 layers of passivator/Seitz-Supradur 100 filter plate composite, a 0.4 μηι filter at the outlet end, and then pumped into passivation with 8% dextran. Hours, dry and disinfected for use. This filter can be used to remove leukocytes and viral inactivating agents such as methylene blue.

Example ^3. Preparation of the device of the invention ( ² )

The apparatus prepared in the embodiment, wherein the solid phase medium comprises the above-mentioned photosensitizer physical adsorption fiber, such as natural fiber, polyamide fiber, polyurethane fiber and filter plate (for example, Supmdur 100, Supradur 500 Eco l000, Permadur 0/400A T2600 described above). , Bio20, Bio40, Bio60, etc.).

The apparatus prepared in this embodiment has the same identification method as the corresponding apparatus prepared in the second embodiment. In general, the apparatus prepared in this example has a similar photosensitizer virus inactivating agent removal ability and has significantly lower side effects than the existing apparatus (for example, a device containing activated carbon or an inorganic adsorption medium). For example, protein adsorption is 15% lower or / and APTT value is lower than 15%), or there is obvious Lower particle contamination (for example, the diameter of the biological fluid flowing through the treatment system visible under the microscope is greater than

Ιμιη has a lower particle count of 15% or more).

The following examples give a few more detailed examples.

Example 3.1 Dye or / and psoralen photosensitizer virus inactivating agent removal filter

The apparatus prepared in this example (including 3.1.1 and 3丄 2) is suitable for the removal of dyes or/and psoralen-type photosensitizer virus inactivating agents. In this embodiment, the photosensitizer virus inactivating agents used are methylene blue, phthalocyanine dye, hypericin, cyanine dye and psoralen.

Example 3.1.1 Virus inactivating agent removal filter containing photosensitizer virus inactivating agent adsorption fiber

In this embodiment, the virus inactivating agent adsorbing fiber is a fiber of an organic polymer physical adsorbent containing a dye or/and a psoralen photosensitizer virus inactivating agent, such as ultrafine polypropylene oil absorbing fiber or/and poly Polyurethane fiber. The fiber loading thickness is 5 cm.

Embodiment 3.1.2 Virus Inactivating Agent Removal Filter Containing Photosensitizer Virus Inactivating Agent Adsorbing Fiber and Adsorbing Particles In this embodiment, the adsorbing particles are dye-containing or/and psoralen-based photosensitizer virus inactivating agents. Granules such as polypropylene matrix resins (e.g., HP2MG series), styrene containing macroporous adsorption resins (e.g., Amberlite XAD-7HP Amberlite XAD-16), and the like.

In this embodiment, the fibers used are contained in the filter material used, such as various Seitz products (KS80, K900, Supradur 80 > P30, Eco 1000, Permadur 0/400A, T2600, Bio20, Bio40, Bio60, Supradur 100, and many more). These filter plates have an adsorption effect on photosensitizer virus inactivating agents, particularly dye-based photosensitizers. The filter has a diameter of 1.5 cm, the macroporous adsorption particles are packed to a thickness of 3 cm, and a 3.5 mm thick filter plate is loaded under the adsorbed particles. In addition to further removing the virus inactivating agent, these filter plates prevent the macroporous adsorption particles from falling off and contaminating the biological fluid to cause side effects.

Example 3.2 Dye-like virus inactivating agent removal filter

A device prepared in accordance with this embodiment (including 3.2.1-3.2.4) is suitable for removing a dye-based photosensitizer virus inactivating agent such as methylene blue.

Example 3.2.1 Virus inactivating agent removal filter containing polyolefin fiber filter plate

In this embodiment, the virus inactivating agent adsorption medium is selected from the group consisting of a polyolefin fiber filter material of an organic polymer physical adsorbent containing a dye-based photosensitizer virus inactivating agent, such as the above-mentioned Eco 1000, Permadur 0/400A, T2600, Supradur. 80> Supradur 100, Supradur 500. The filter plate is filled with more than 3 layers (each thickness is 3.5mm). Example 3.2.2 Virus inactivating agent removal filter containing natural fiber filter plate

In this embodiment, the virus inactivating agent adsorption medium is selected from the group consisting of natural fiber-containing filter materials, such as various Seitz company products (e.g., Bio20, Bio40, Bio60, etc.). The filter plate is filled with more than 3 layers (each thickness is 3.5mm).

Example 3.2.3 Virus inactivating agent removal filter containing natural fiber In this embodiment, the virus inactivating agent adsorption medium is selected from natural fibers such as cotton wool fibers, silk fibers or wood fibers.

Example 3.2.4 Virus inactivating agent removal filter containing synthetic fiber

In this embodiment, the virus inactivating agent adsorption medium is selected from the group consisting of synthetic fibers containing organic polymer physical adsorbents, such as polyurethane fibers or polystyrene fibers.

Example 3.3 Dye-like virus inactivating agent removal / removal of leukocyte filter

Some of the devices prepared as described above in Example 3 can also be used for dye virus inactivating agents to remove and remove leukocytes. In fact, cotton fibers, polyesters, polyurethanes, and the like, in this embodiment, are both a dye-based photosensitizer virus inactivating agent adsorption medium and a leukocyte-removing material.

Example 4. Preparation of the device of the present invention

The apparatus prepared in this embodiment, wherein the solid phase medium contains the above organic solvent physical adsorption medium. The apparatus prepared in this example was identified in the same manner as the apparatus prepared in Example 2. In general, the devices prepared in this example have similar virus inactivating agent removal/and virus inactivation efficiencies compared to existing devices, with significantly lower side effects (eg, with activated carbon or inorganic adsorption). Comparison of medium devices): Protein adsorption is 15% lower or / and APTT value is lower than 15%.

The following examples give a few more detailed examples.

Example 4.1 Organic solvent virus inactivating agent removal device

The apparatus of this embodiment is prepared by charging the above organic solvent physical adsorption medium into a filter drum or a hollow column. Organic solvent physical adsorption medium used, including fibers (for example, ultrafine polypropylene oil absorbing fibers), filter plates (for example) or/and macroporous adsorption resins (such as Supradur 80), polyphenylene-containing porous particles (Amberlite XAD-7HP) AmberliteXAD-16, etc.).

Example 4.2 Organic solvent virus inactivation device

The apparatus of the present embodiment, a preparation method, is prepared by charging an organic solvent virus inactivating medium into a filter drum or a hollow column. In the present embodiment, the organic solvent virus inactivating medium contains the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium. The preparation method used in this method is the same as the method of "fixing the virus inactivating agent to the adsorption medium" in Example 1.2, wherein: the adsorption medium used is selected from the above organic solvent physical adsorption medium (for example, used in Example 4.1). The organic solvent physical adsorption medium); the virus inactivating agent used is S (for example, TnBP or β-propiolactone) or SD (for example, ΤηΒΡ/earth temperature-80, TnBP/Triton X100, or TnBP/alcohol).

Another preparation method is: loading the above organic solvent physical adsorption medium into a filter drum or a hollow column, pre-washing, and then using an organic solvent dispersion system to flow through the adsorption medium at an optimized speed (for example, 0.1-lcm/min) to remove the organic solvent. The virus inactivating agent is fixed and then the unfixed material is washed out. The preparation method of the organic solvent dispersion system is the same as the method of "preparation of the organic solvent dispersion system" in Example 1.1. Organic solvent Examples of the dispersion system include the following solutions in appropriate concentration ratios: TnBP/earth temperature-80 solution, TnBP/Triton 5Q00 solution, TnBP/alcohol solution, and the like.

The content of the organic solvent-inactivated medium containing the organic solvent inactivating agent may vary depending on the needs. For example, if the volume of the biological fluid to be treated is 250 ml, the TnBP content of some devices is greater than 7.5 mg, and the TnBP content of some devices is equal to or less than 7.5 mg, or even equal to or less than 2.5 mg. It corresponds to the content of the organic solvent virus inactivating agent respectively greater than 3 X (lOppmX volume of the single-part blood component ml), equal to or less than 3 X (lOppmX volume of the single-part blood component ml), And equal to or less than (lOppmX the volume of the single-part blood component ml).

Example 4.3 Organic solvent virus inactivation / organic solvent virus inactivating agent removal device

The apparatus of this embodiment is prepared by separately loading an organic solvent virus inactivating medium and an organic solvent virus inactivating agent adsorption medium into a filter drum or a hollow column. There are two filling methods: A). The organic solvent virus inactivation medium is placed near the inlet of the chamber, and the organic solvent virus inactivating agent adsorption medium is placed near the outlet; B) The organic solvent virus inactivating agent is adsorbed The medium is placed near the inlet of the chamber and near the outlet, and the organic solvent virus inactivating medium is placed therebetween. Alternatively, the organic solvent virus inactivating device may be connected in series with the organic solvent virus inactivating agent removing device.

In this embodiment, the solid phase medium used is a combination of one of the following: 1) an organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (refer to the implementation) Example 4.2), and the above organic solvent physical adsorption medium (refer to Example 4.1); 2). Other organic solvent virus inactivating medium (for example, the organic solvent virus inactivating medium prepared in Example 1), and the above organic solvent physical adsorption medium (Reference Example 4.1); 3). Organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and organic solvent virus inactivating agent immobilized on the adsorption medium (refer to Example 4.2), and other organic solvent viruses The active agent adsorption medium (for example, the organic solvent virus inactivating agent adsorption medium prepared in Example 1).

EXAMPLES 4.4 Organic solvent virus inactivation / Organic solvent virus inactivating agent removal / Photosensitizer virus inactivating agent removal device

The apparatus of the present embodiment is an organic solvent virus inactivating medium, an organic solvent virus inactivating agent adsorption medium, and a photosensitizer virus inactivating agent adsorption medium (or an organic solvent virus inactivating medium and an organic solvent virus inactivating agent/photosensitive). The agent virus inactivating agent adsorption medium is prepared by separately loading into a filter drum or a hollow column. There are two methods of filling: A). The organic solvent virus inactivating medium is placed near the inlet of the chamber; B) The organic solvent virus inactivating agent medium is placed near the middle of the chamber. Alternatively, it may be formed by an organic solvent virus inactivating device, an organic solvent virus inactivating agent removing device, and a photosensitizer virus inactivating agent removing device.

In this embodiment, the solid phase medium used is a combination of one of the following: 1) an organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (see Test Example 4.2), the above organic solvent physical adsorption medium (refer to Example 4.1), and the above photosensitizer physical adsorption fiber (Reference Example 3); 2). Other organic solvent virus inactivation medium (for example, prepared in Example 1) Organic solvent virus inactivating medium), the above organic solvent physical adsorption medium (refer to Example 4.1), and the above photosensitizer physical adsorption fiber (refer to Example 3), or other photosensitizer adsorption medium (for example, the photosensitizer prepared in Example 1) Virus inactivating agent adsorption medium); 3). Organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and organic solvent virus inactivating agent immobilized on the adsorption medium (refer to Example 4.2), and other organic solvent viruses Inactivator adsorption medium (for example, organic solvent virus inactivating agent adsorption medium prepared in Example 1), and photosensitizer adsorption medium (for example, the above photosensitizer physical adsorption fiber or the photosensitizer virus inactivating agent adsorption medium prepared in Example 1) 4). An organic solvent virus inactivating medium containing the above organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized on the adsorption medium (Reference Example 4.2) And a photosensitizer / organic solvent virus inactivating agent adsorption medium (e.g., a polymer containing styrene macroporous adsorption resin, passivating agent prepared in Example 1 / activated carbon composite, etc.).

Example 5. Preparation of the device of the present invention (4)

The apparatus prepared in this example, wherein the solid phase medium is a combination of solid phase media in different apparatuses prepared in the above examples.

When a plurality of solid phase media are combined, there are a series relationship or a parallel relationship between different solid phase media. In this example, only some examples of solid-phase media in series are listed: Combination of adsorbent/passivator complex with virus inactivating media, adsorbent/passivator complex and adsorbent/passivator complex Combination, adsorption medium/passivator composite and fiber filter; The serial combination of solid phase media can be cascaded in the same container or in multiple vessels. In this case, use the method of concatenation in the same container. Table 3 lists some of the devices prepared in this example.

table 3

Device solid phase medium 1 solid phase medium 2 function

Filter F1 A9 (Table 1) Seitz- Bio 20 In addition to SD agent / photosensitizer virus inactivating agent and its derivatives

Filter F2 A9 (Table 1) Seitz-Supradur In addition to SD agent / photosensitizer virus inactivating agent

100 derivatives

Filter F3 A9 (Table 1) Seitz-T2600 In addition to SD agent / photosensitizer virus inactivating agent and its derivatives

Filter F4 A9 (Table 1) A12 (Table 1) Except for SD agent / photosensitizer virus inactivating agent and its derivatives

Filter F5 A12 (Table 1) A21 (Table 1) Except for SD agent / photosensitizer virus inactivating agent and its derivatives

Filter F6 A15 (Table 1) A28 (Table 1) In addition to SD agent / photosensitizer virus inactivating agent and its derivatives Filter F7 A20 (Table 1) A28 (Table 1) In addition to SD agent / photosensitizer virus inactivating agent and its derivatives

Filter F8 A23 (Table 1) A24 (Table 1) Except for SD agent

Filter F9 A9 (Table 1) Seitz- Bio 40, in addition to SD agent / photosensitizer virus inactivating agent and its polyester oil-absorbing fiber derivative / leukocyte

Filter G1 B6 (Table 2) B 12 (Table 2) Double virus inactivation

Filter G2 B6 (Table 2) B 14 (Table 2) Double virus inactivation

Filter G4 B6 (Table 2) A23 (Table 1) Solid phase SD virus inactivation

Filter G5 B 12 (Table 2) A21 (Table 1) Solid phase SD virus inactivating agent virus inactivation

Filter G6 B 13 (Table 2) A29 (Table 1) Solid phase iodine virus inactivation

Filter G7 C2 Seitz- Bio 20, solid phase SD virus inactivating agent virus inactivation

Propylene oil absorbing fiber

Filter G8 C2 Seitz- Bio 40, polymeric phase SD virus inactivating agent virus inactivation / de-white lipid absorption fiber

Filter K1 S/D-PS Amino acid passivated solid phase SD virus inactivation

With resin PS adsorption resin

The function and side effects of the filter in this embodiment are consistent with the functions and side effects of the solid phase medium contained therein.

(Refer to the related embodiment above).

Example 6. Example of a single blood component kit of the present invention

In the present invention, the term "single-part blood component processing device" refers to a biological liquid processing device in which a single-part blood component is a biological liquid. The single-part blood component processing device of the present invention includes single-part plasma Virus inactivation device, single-part platelet virus inactivation device (for example, using psoralen as a virus inactivating agent), etc. Based on the above device, several other devices may also be derived. For example, by interacting with other processing systems (eg, The blood collection and separation processing system are connected to form a more integrated device.

The kit prepared in this embodiment (including 6.1 to 6.3) contains at least a single blood component treatment device, and is a device prepared in the above embodiment.

In some cases, the kit also contains a viral inactivating agent addition structure, including a liquid phase virus inactivating agent addition structure or a solid phase virus inactivating agent addition structure. Examples of liquid phase viral inactivating agents (eg, photosensitizer virus inactivating agent solutions) added to the structure are: medical polystyrene plastic tubes, liquid phase virus inactivating agents in tubes, and deblockable closed structures (eg, switches or Fragile medical polystyrene plastic sheet).

In some cases, the kit also contains a viral inactivation liquid reaction site, such as a clear plastic bag for the photosensitizer virus inactivation, and the like.

In some cases, the kit also contains other structures (eg, in addition to leukocyte filters, lotions, lotion addition devices, etc.), where a lotion (eg, saline) is used to place the device, particularly a column device or / The blood components retained on the solid phase medium in the filter device are washed out to reduce loss of blood components.

The function and side effects of the kit in this embodiment are consistent with the functions and side effects of the device contained therein (refer to the related embodiments above). If other parts of the kit have unwanted reactivity, follow the instructions. It is necessary to pass a substance selected from the above passivating agent for passivation. The passivation methods include: 1) passivating and reassembling the parts that need to be passivated into devices; 2) passivating the assembled device; 3) passivating and reassembling the parts that need to be passivated The device is then re-passivated for the assembled device.

Example 6.1 Single Human Blood Component Virus Inactivation Kit

Example 6.1.1 Single Human Blood Component Virus Inactivating Kit Containing Virus Inactivating Agent Adsorption Treatment System

The kit prepared in this embodiment comprises a virus inactivating agent adsorption treatment system, i.e., a filter or column containing the virus inactivating agent adsorption medium prepared in the above embodiment. The kit also contains a viral inactivating agent addition device and a virus inactivation reaction site. The working principle is as follows: the single blood component is contacted with the virus inactivating agent added by the virus inactivating agent, and the virus inactivation reaction is carried out at the virus inactivation reaction site, and the biological fluid flows at a preferred flow rate after the reaction is completed. The virus inactivating agent adsorbs the filter or column of the medium and removes the virus inactivating agent or/and its derivative, and if necessary, washes the retained blood component by adding the washing liquid to the structure. Example 6.1.2 Single Human Blood Component Virus Inactivating Kit Containing Virus Inactivation Treatment System

The kit prepared in this embodiment comprises a virus inactivation treatment system, i.e., a virus inactivating medium-containing filter or column prepared in the above embodiment. The working principle is as follows: a single human blood component flows through a filter or column containing a virus inactivating medium at a preferred flow rate for virus inactivation (for example, immobilized SD agent treatment, immobilized iodine treatment, or immobilized ion treatment) If necessary, the blood components retained in the treatment system can be washed out by adding the washing liquid to the structure and adding the washing liquid.

Example 6.1.3 Single-part blood component virus inactivating kit containing virus inactivation/viral inactivating agent adsorption treatment system

The kit prepared in this embodiment comprises a virus inactivation treatment system, that is, a filter or column containing the virus inactivating medium and the virus inactivating agent adsorption medium prepared in the above embodiment. The working principle is as follows: The single blood component is subjected to virus inactivation by a preferred flow rate filter or a virus inactivating medium in the column (for example, immobilized SD treatment, immobilized iodine treatment, or immobilized ion treatment: And flowing through the virus inactivating agent adsorption medium to ensure that the blood component of the effluent filter or column is substantially free of viral inactivating agents. If necessary, the blood components retained in the treatment system can be washed out by adding the washing liquid to the structure.

Example 6.1.4 Single Human Blood Component Virus Inactivating Kit Containing Dual Virus Inactivation Treatment System

The kit prepared in the present embodiment, which comprises the apparatus prepared in Example 5, further comprises a virus inactivating agent addition device and a first virus inactivation reaction site. The working principle is as follows: the single blood component is contacted with the virus inactivating agent added by the virus inactivating agent, and the virus inactivation reaction is carried out at the virus inactivation reaction site, and the biological liquid is flowed through at a preferred flow rate after the reaction is completed. The filter or column prepared in Example 5 was subjected to virus inactivation and virus inactivating agent removal, and the blood components exiting the filter or column were substantially free of viral inactivating agents. If necessary, the blood components retained in the treatment system can also be washed out by adding the washing liquid to the structure to add the washing liquid. Example 6.2 Single Human Blood Component Virus Inactivation/De-Leukocyte Kit The kit prepared in this embodiment, comprising a leukocyte-removing and virus inactivating agent adsorption treatment system, that is, the de-leukocyte-containing and virus inactivating agent adsorption medium prepared by the above embodiment (or containing de-white blood cells and virus inactivating medium, or A filter or column that removes white blood cells and virus inactivating media and virus inactivating media.

Example 7. Virus inactivation method of the present invention (1)

The method of the present embodiment (including 7.1-7.4), comprising at least: A) providing a biological fluid; B) providing a processing system, the processing system comprising at least a solid phase medium and a chamber containing the solid phase medium, and At least a portion of the solid phase medium comprises a passivating adsorption medium; C). flowing the biological fluid through the processing system for viral inactivation or viral inactivating agent adsorption. The treatment system used in this example is selected from the corresponding apparatus comprising the passivated adsorption medium prepared in Example 2, 5 or 6.

EXAMPLES 7.1 Method for adsorption removal treatment of virus inactivating agent

Example 7.1.1 Example of inactivation of virus by single blood component

In this embodiment, the biological fluid used is a single-part platelet; the virus inactivating agent used is psoralen (4,-aminomethyl-4,5,8-trimethyl psoralen); a kit selected from the preparation of Example 6, which comprises the device prepared in Example 2.1, which comprises a psoralen-type virus inactivating agent adsorption medium combined with a passivating agent (for example, polyester oil absorbing fiber, glass fiber and super Fine polypropylene oil absorbing fiber).

In this embodiment, the virus inactivation method is: adding the virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing virus inactivation at the virus inactivation reaction site (appropriate concentration of psoralen, room temperature, illumination 30) Minutes), then the biological fluid is introduced into the device and the virus inactivating agent is removed by contact with the virus inactivating agent adsorption medium therein.

The method of this example is also effective in removing viral inactivating agents (e.g., more than 90% of psoralen removal) with a smaller side effect, as compared to a method using a similar device containing a conventional adsorption medium (e.g., activated carbon). For example, in a control method using a device containing activated carbon felt, the protein loss was 10-20%, the platelet loss was 7%, and the platelet morphology score was decreased by 16%. However, using the adsorption filter prepared in 2.1, the protein loss was only 5%. The platelet loss was 3%, and the platelet morphology score did not decrease.

Example 7.1.2 Virus inactivation of other biological fluids

In this embodiment, the biological fluids used are: 1. multi-part combined plasma, 2. human plasma fraction containing human fibrinogen (for example, component I or cryoprecipitate in cold alcohol precipitation), 3. Human plasma separation component (eg, cryoprecipitate) containing human coagulation factor VIII, 4, human plasma separation component containing human coagulation factor IX (eg, DEAE-Sephadex washout component of cryoprecipitate supernatant), 5. Human plasma separation component of human prothrombin complex (PCC) (eg DEAE-Sephadex washout component of cryoprecipitate supernatant), 6. Human plasma fraction containing gamma globulin (eg cold alcohol precipitation method) Component II precipitation:), 7. Recombinant Alpha interferon (interferon concentration 1.15 million IU/ml, pH 7.0), 8. Positive human serum reference.

In this embodiment, the virus inactivating agent used is an S/D agent (for example, TnBP/Triton X100, TnBP/spit Temperature 80, or diethyl ether / Triton X100); the apparatus used is selected from the adsorption filter prepared in Example 2.1 (the size of which varies depending on the amount of biological liquid), and the adsorption filter contains an organic solvent virus with a passivating agent immobilized thereon. Living agent adsorption medium (for example, styrene-containing polymer macroporous adsorption resin or activated carbon filter plate).

The virus inactivation method in this embodiment is: adding the virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing virus inactivation at the virus inactivation reaction site (0.3-1% S, or 0.3-1%) S and 0.3-1% D; 30 ° C; 4 hours), then the biological liquid is introduced into the above device, and the organic solvent virus inactivating agent is removed by contact with the virus inactivating agent adsorption medium therein.

The method of this example is also effective in removing viral inactivating agents (e.g., more than 90% of TnBP removal) compared to methods using similar devices containing conventional adsorption media (e.g., activated carbon), with side effects being much smaller. For example, the virus inactivation method in this example results in an increase in plasma APTT value of less than 30% compared to existing methods using activated carbon, such that the biological activity of other biological fluids (eg, fibrinogen, coagulation factor VIII) The loss rate of coagulation factor IX and total protein is 30% or less.

EXAMPLES 7.2 Method for inactivating treatment with immobilized virus inactivating agent virus

Example 7.2.1 Treatment of single blood component

In the present embodiment, the biological fluid used is a single-part plasma; the apparatus used is selected from the kit prepared in Example 6, which comprises the virus inactivating apparatus prepared in Example 2.2, which respectively contains the organic solvent virus inactivating agent/adsorption medium. / Passivator complex, Crnio-Zetaplus VR virus filter plate and Seitz-iodine-PVPP filter plate. It should be emphasized that the virus inactivation treatment in the present embodiment can enable the plasma product factory to perform virus inactivation after the pulping to reduce the risk of virus contamination in the blood storage, transportation and production process, and improve the product. Virus security.

The virus inactivation method in this embodiment is: a biological fluid at room temperature is passed through the above apparatus at a preferred flow rate (e.g., a linear velocity of 0.1 - 1.5 cm / min:) to carry out virus inactivation. If necessary, wash the device including the solid phase media with sputum.

Compared with the method using a similar device containing a common adsorption medium (for example, activated carbon), the method of the present embodiment can equally effectively inactivate the virus (for example, the pattern virus is inactivated by more than 4 logs), and has less unnecessary Reactivity (for example, the APTT value is increased by more than 20%, and the loss rate of fibrinogen, coagulation factor VIII, coagulation factor IX, and total protein is 15% or less).

Example 7.2.2 Treatment of other biological fluids

In this embodiment, the biological fluids used are: 1. multi-part combined plasma, 2. human plasma fraction containing human fibrinogen (for example, component I or cryoprecipitate in cold alcohol precipitation), 3 Human plasma separation component of human prothrombin complex (PCC) (eg, DEAE-Sephadex washout component of cryoprecipitate supernatant). In this embodiment, the apparatus used is selected from the virus inactivation apparatus prepared in Example 2.2 (the volume of the chamber is different from the volume of the biological liquid), and the adsorbent medium/organic solvent virus inactivating agent/passivator complex is respectively contained. , Cuno-Zetaplus VR virus-killing filter plate and Seitz-iodine-PVPP filter plate.

The virus inactivation method in this example is the same as in the method of Example 7.2.1, and the comparison with the method using a similar apparatus containing a usual adsorption medium (e.g., activated carbon) is also in agreement with the comparison in Example 7.2.1. EXAMPLE 7.3 Method for removing steps of virus inactivating agent and leukocyte

In this embodiment, the biological fluid used is a single-part plasma; the virus inactivating agent used is selected from the photosensitizer virus inactivating agent (for example, 4,-aminomethyl-4,5,8-trimethyl psoralen) The kit used was selected from the kit prepared in Example 7, which contained the leukocyte/virus inactivating agent removal filter prepared in Example 2.4.

The leukocyte removal/viral inactivating agent removal method in this embodiment is: adding a virus inactivating agent to a biological liquid through a virus inactivating agent addition device, and performing virus inactivation at a virus inactivation reaction site (for example, an appropriate concentration of virus) The inactivating agent, room temperature, light for 30 minutes), then the biological liquid is introduced into the above device, and the virus inactivating agent and its derivative are removed by contact with the solid phase medium therein.

The method of this example has an equivalent leukocyte removal efficiency and a significant reduction in side effects as compared to a method using a similar device containing a conventional adsorption medium such as glass fiber. Example 8. Virus inactivation method of the present invention (2)

The method of the present embodiment (including 8.1-8.3), comprising at least: A) providing a biological fluid; B) contacting the biological fluid with a viral inactivating agent and performing virus inactivation (eg, an appropriate concentration of virus is eliminated) Living agent, room temperature, illumination for 30 minutes), wherein the virus inactivating agent comprises a photosensitizer virus inactivating agent; C) providing a treatment system, the treatment system containing at least a virus inactivating agent adsorption medium and containing the solid phase a chamber of the medium, wherein the virus inactivating agent adsorption medium contains a photosensitizer physical adsorption fiber; D). flowing the biological liquid treated by the step B) through the treatment system to remove the photosensitizer virus inactivating agent adsorption and its derivative .

In the present embodiment, the biological fluid used was a single-part plasma; the treatment system used was the apparatus prepared in Example 7, which contained a device (filter or column) selected from the preparation of Example 3.

The method of the present embodiment can effectively remove inactivated virus agents (e.g., more than 95%) with less unwanted reactivity (e.g., compared to methods using similar devices containing conventional adsorption media (e.g., activated carbon). The APTT value is increased by more than 20%, and the loss rate of fibrinogen, coagulation factor VIII, coagulation factor IX, and total protein is 15% or less).

EXAMPLES 8.1 Method for removing a dye-containing or/and psoralen-based photosensitizer virus inactivating agent

In this embodiment, the virus inactivating agent used is selected from the group consisting of dyes or psoralen photosensitizer virus inactivating agents (for example, methylene blue or 4,-aminomethyl-4,5,8-trimethyl) Psoralen:) The device used was selected from the kit prepared in Example 6, which contained the adsorption filter prepared in Example 3.1.

EXAMPLES 8.2 Method for removing step of dye-containing photosensitizer virus inactivating agent

In this embodiment, the virus inactivating agent used is selected from the group consisting of a dye-based photosensitizer virus inactivating agent (for example, methylene blue); The apparatus used was selected from the kit prepared in Example 6, which contained the adsorption filter prepared in Example 3.2.

EXAMPLES 8.3 Method for removing/removing leukocyte-containing step of dye-containing photosensitizer virus inactivating agent

In the present embodiment, the virus inactivating agent used is selected from a dye-based photosensitizer virus inactivating agent (e.g., methylene blue); and the apparatus used is selected from the kit prepared in Example 6, which contains the adsorption filter prepared in Example 3.3.

Example 9. Virus inactivation method of the present invention (3)

The virus inactivation method in this embodiment (including 9.1-9.3) includes at least: A) providing a biological fluid; B) providing a treatment system, the treatment system comprising at least a solid phase medium and a chamber containing the solid phase medium Wherein the solid phase medium comprises a virus inactivating medium or/and a virus inactivating agent adsorption medium, and: the virus inactivating medium comprises at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon, The virus inactivating agent adsorption medium comprises at least an organic solvent physical adsorption medium; C). flowing the biological liquid through the treatment system for virus inactivation or/and virus inactivating agent adsorption. The treatment system used in this example was selected from the apparatus prepared in Example 4.

EXAMPLES 9.1 Method for adsorption removal step of organic solvent-containing virus inactivating agent

Example 9.1.1 Example of inactivation of virus by single blood component

In the present embodiment, the biological fluid used was a single-part plasma; the apparatus used was selected from the kit prepared in Example 6, which contained the apparatus prepared in Example 4.1.

The virus inactivation method in this example is the same as the virus inactivation method in Example 7.1.1, and the virus inactivating agent removal efficiency and side effect reduction are also the same.

Example 9.1.2 Examples of virus inactivation of other biological fluids

In the present embodiment, the biological fluid used is the same as that used in the embodiment 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.1 (the volume of the chamber varies depending on the volume of the biological fluid). The virus inactivation method in this embodiment is the same as the virus inactivation method in Example 7.1.2, and the virus inactivating agent removal efficiency and side effect reduction are also the same.

EXAMPLES 9.2 Method for Inactivating Step of Immobilized Organic Solvent Virus

Example 9.2.1 Example of inactivation of virus by single blood component

In the present embodiment, the biological fluid used was a single-part plasma; the apparatus used was selected from the kit prepared in Example 6, which contained the apparatus prepared in Example 4.2. The virus inactivation method in this example is the same as the virus inactivation method in Example 7.2.1, and the virus inactivation efficiency and side effect reduction are also the same.

Example 9.2.2 Examples of virus inactivation of other biological fluids

In the present embodiment, the biological fluid used is the same as the biological fluid used in Example 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.2 (the volume of the chamber is different from the volume of the biological fluid). The virus inactivation method in this embodiment is the same as the virus inactivation method in Example 7.2.2, and the virus inactivation efficiency and side effect reduction are also the same. Example 9.3 Method for Removal of Immobilized Organic Solvent Virus Inactivation/Organic Solvent Virus Inactivating Agent Example 9.3.1 Single Human Blood Component Virus Inactivation Example

In the present embodiment, the biological fluid used is a single-part plasma; the apparatus used is selected from the kit prepared in Example 6, which contains the filter or column prepared in Example 4.3.

The virus inactivation method in this embodiment is: flowing the biological fluid at room temperature through the above device at a preferred flow rate, performing virus inactivation and removing the organic solvent virus inactivating agent which may fall off.

The virus inactivation efficiency and side effects in the method of this example were the same as in the method of Example 9.2.2, and the content of the organic solvent in the biological liquid collected from the outlet of the apparatus used was less than 10 ppm.

Example 9.3.2 Examples of virus inactivation of other biological fluids

In the present embodiment, the biological fluid used is the same as that used in the embodiment 7.2.2; the apparatus used is selected from the filter or column prepared in Example 4.3 (the volume of the chamber varies depending on the volume of the biological fluid).

The virus inactivation method in this embodiment is: flowing the biological fluid at room temperature through the above device at a preferred flow rate, performing virus inactivation and removing the organic solvent virus inactivating agent which may fall off. The virus inactivation efficiency, side effects, and organic solvent content of the collected biological fluids in the method of this example were consistent with the method of Example 9.3.1.

Example 10. Viral inactivation method of the present invention (4)

The method of virus inactivation in this embodiment (including 10.1 and 10.2) comprises at least: A) providing a biological fluid; B) performing a first viral inactivation of the biological fluid; C) providing a treatment system, The treatment system comprises at least a virus inactivating medium and a chamber containing the solid phase medium, and wherein the virus inactivating medium contains at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon; D). The biological fluid inactivated by the first virus flows through the treatment system for a second viral inactivation. In this embodiment, the biological fluid used is a single blood paddle. In this embodiment, the apparatus used is selected from the kit prepared in Example 6, which comprises a device selected from the preparation of Example 4.4 or Example 5.

In this embodiment, two different virus inactivation mechanisms are used, and the virus inactivation efficiency is high.

EXAMPLES 10.1 Virus Inactivation of Photosensitizer Virus Inactivation and Organic Solvent Virus Inactivation Steps

In this embodiment, the photosensitizer virus inactivating agent used is selected from the group consisting of dyes or psoralen photosensitizer virus inactivating agents (for example, methylene blue or 4'-aminomethyl-4, 5, 8-3 Methyl psoralen). In this embodiment, the treatment system used is selected from the apparatus prepared in Example 4.4 or Example 5, in particular, a photosensitizer virus inactivating agent adsorption medium, an organic solvent virus inactivating medium, and an organic solvent virus inactivating agent adsorption medium. Wherein the virus inactivating medium comprises at least an organic solvent physical adsorption medium and an organic solvent virus inactivating agent immobilized thereon.

In this embodiment, the virus inactivation method is: adding the photosensitizer virus inactivating agent to the biological liquid through the virus inactivating agent adding device, and performing the first virus inactivation at the virus inactivation reaction site (for example, an appropriate concentration) Virus inactivating agent, room temperature, light for 30 minutes), then let the biological fluid enter the above device, remove the photosensitizer virus inactivating agent by contact with the solid phase medium therein and carry out the second virus inactivation, and remove the photosensitizer virus Inactivating agent and organic solvent virus inactivating agent.

In this embodiment, the side effects are controlled at an acceptable level due to the use of a solid phase medium having a lower side effect. For example, after the single-person plasma is inactivated by the above virus, the APTT value is kept within 47 seconds, and the loss rates of fibrinogen, coagulation factor VIII, and coagulation factor IX are also less than 30%, respectively.

Example 10.2 Virus inactivation with virus heat inactivation and organic solvent virus inactivation steps

In the present embodiment, the apparatus used is selected from the kit prepared in Example 6, which comprises the filter or column prepared in Example 2.2 or 4.3, which contains an organic solvent virus inactivating medium and an organic solvent virus inactivating agent adsorption medium.

The method for inactivating the virus in this embodiment is: adding a known plasma active protective agent (for example, sugar, sodium valerate, calcium, etc.) to the biological liquid through the protective agent adding device, and performing virus at the virus inactivation reaction site. Inactivate (for example, a suitable concentration of protective agent, 52 ° C, 3 hours), then let the biological liquid enter the above device, inactivate the organic solvent virus by contact with the solid phase medium and remove the organic solvent that may fall off Active agent.

In this embodiment, the side effect of at least the second virus inactivation is small due to the use of a solid phase medium having a lower side effect.

Example 11. Virus inactivation method of the present invention (5)

The virus inactivation method in this embodiment at least comprises: A) providing a biological fluid; B) providing the device prepared in Example 2, comprising the virus inactivating medium or/and a virus inactivating agent adsorption medium; The biological fluid is passed through the treatment system for virus inactivation or/and viral inactivating agent adsorption, and leukocyte removal.

It will be apparent that various modifications and changes can be made to the preferred embodiments of the invention described herein. These modifications and variations can be made without departing from the spirit and scope of the invention and the advantages thereof. Accordingly, such modifications and variations are intended to be included within the scope of the appended claims.

Claims

Claim

A method for inactivating a virus, comprising at least:

A) providing a biological fluid; B) providing a treatment system; and C) contacting the biological fluid with the treatment system,

Wherein the processing system comprises at least a solid phase medium and a chamber containing the solid phase medium,

Wherein the solid phase medium comprises at least:

a) a virus inactivating agent adsorption medium; and / or

b) a virus inactivating medium comprising a virus inactivating agent adsorption medium and a virus inactivating agent immobilized thereon,

Wherein the virus inactivating agent adsorption medium comprises:

(a) passivating an adsorption medium containing at least an adsorption medium and a passivating agent immobilized on the adsorption medium; or / and

(b) physical adsorption medium, the physical adsorption medium comprising:

(i) a photosensitizer physically adsorbing fiber used as a photosensitizer virus inactivating agent adsorption medium, said photosensitive agent physically adsorbing an organic polymer physical adsorbent containing a photosensitizer virus inactivating agent;

(ii) An organic solvent physical adsorption medium used as an organic solvent virus inactivating agent adsorption medium, the organic solvent physical adsorption medium containing an organic solvent physical adsorbent of an organic solvent virus inactivating agent.

2. The method of claim 1, wherein the solid phase medium comprises an organic solvent virus inactivating agent adsorption medium, and the organic solvent virus inactivating agent adsorption medium contains at least the passivation adsorption medium, or/and organic Solvent physical adsorption medium.

3. The method of claim 1, wherein the solid phase medium comprises an organic solvent virus inactivating medium, and the organic solvent virus inactivating medium comprises at least an organic solvent virus inactivating agent, and the passivating adsorption medium, Or / and organic solvent physical adsorption medium.

The method of any one of claims 1 to 3, wherein the organic solvent physic adsorption medium comprises macroporous resin or/and fiber.

The method according to any one of claims 1 to 4, wherein the organic polymer physical adsorbent of the organic solvent virus inactivating agent comprises a polyolefin.

6. The method of claim 5 wherein the polyolefin comprises polystyrene.

7. The method of claim 1 wherein said solid phase medium comprises a photosensitizer virus inactivating agent adsorption medium, and said photosensitizer virus inactivating agent adsorption medium comprises at least said passivated adsorption medium, or/and photosensitive The agent physically adsorbs the fiber.

8. The method of any of claims 1 or 7, wherein the photosensitizer physico-adsorbing fibers comprise natural fibers.

9. The method of claim 8 wherein the natural fibers comprise wood fibers.

10. The method of claim 1, 7 or 8, wherein: said photosensitizer physically adsorbing fibers comprise synthetic fibers.

11. The method of claim 10, wherein the synthetic fibers comprise polyolefin fibers.

The method of any one of claims 1 to 10, wherein the solid phase medium comprises the organic solvent virus inactivation medium, the organic solvent virus inactivating agent adsorption medium, or/and the photosensitizer physical adsorption fiber.

13. The method of claim 1 wherein said solid phase medium comprises an iodine virus inactivating medium, said iodine virus inactivating medium comprising at least iodine, an iodine adsorption medium and a passivating agent.

The method according to any one of claims 1 to 13, wherein the solid phase medium comprises a depth filter medium, the deep filter medium contains the virus inactivating agent adsorption medium, and has the following characteristics: A) The average density is greater than 0.25 g/cm ³ ; and B). The ash content is less than 1%.

The method of any one of claims 1-14, wherein the adsorbent medium in the passivating adsorption medium comprises activated carbon.

16. The method of any of claims 1-14, wherein the adsorbent medium in the passivating adsorption medium comprises an ion exchange adsorption medium.

The method of any one of claims 1-14, wherein the adsorbent medium in the passivating adsorption medium comprises the physical adsorption medium.

The method of any one of claims 1 to 17, wherein the passivating agent in the passivating adsorption medium comprises a hydrophilic group or/and an oleophilic group, and the solid phase medium pair can be lowered An organic substance that describes the side effects of biological fluids.

19. The method of claim 18, wherein the purifying agent comprises a natural fat.

20. The method of claim 18, wherein the purification agent comprises a hydroxy compound.

21. The method of claim 18, wherein the purification agent comprises an amino acid.

22. The method of claim 18, wherein the purification agent comprises an organic solvent.

The method according to claim 22, wherein the virus inactivating medium is immobilized with an organic solvent virus inactivating agent and an organic solvent deactivator, and the total content of the organic solvent deactivator and the organic solvent virus inactivating agent is More than 0.2 mol/cm ³ , preferably more than 0.3 mol/cm ³ .

24. The method of any of claims 1-23, wherein the treatment system further comprises a leukocyte-free solid phase medium.

25. The method of claim 24, wherein the leukocyte-free solid phase medium comprises the virus inactivating agent adsorption medium.

The method of any one of claims 1 to 25, wherein the biological fluid comprises a single human blood component.

The method according to claim 26, wherein the content of the organic solvent virus inactivating agent in the organic solvent virus inactivating medium is equal to or less than 3 X (10 ppm of the volume of the single-part blood component ml), or Less than (lOppmX the volume of the single-part blood component ml).

28. A treatment system effective to reduce the risk of a virus, which is the treatment system described in the virus inactivation method of any one of claims 1-27.

29. Apparatus for effectively reducing the risk of a virus, comprising at least the processing system of claim 28.

30. The device of claim 29 which is a single human blood component treatment kit.