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WO2019206284A1 - 磁珠纯化系统 - Google Patents

磁珠纯化系统 Download PDF

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Publication number
WO2019206284A1
WO2019206284A1 PCT/CN2019/084590 CN2019084590W WO2019206284A1 WO 2019206284 A1 WO2019206284 A1 WO 2019206284A1 CN 2019084590 W CN2019084590 W CN 2019084590W WO 2019206284 A1 WO2019206284 A1 WO 2019206284A1
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WO
WIPO (PCT)
Prior art keywords
magnetic
purification
magnetic bead
needle
bead purification
Prior art date
Application number
PCT/CN2019/084590
Other languages
English (en)
French (fr)
Inventor
朱金鑫
贺瑞娜
钱红
白涛
李德铭
郑成
陈国栋
Original Assignee
南京金斯瑞生物科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 南京金斯瑞生物科技有限公司 filed Critical 南京金斯瑞生物科技有限公司
Priority to JP2020560367A priority Critical patent/JP7298102B2/ja
Priority to KR1020207033461A priority patent/KR102740873B1/ko
Priority to US17/051,119 priority patent/US12226784B2/en
Priority to SG11202010391WA priority patent/SG11202010391WA/en
Priority to EP19791628.1A priority patent/EP3789478A4/en
Publication of WO2019206284A1 publication Critical patent/WO2019206284A1/zh

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/02—Magnetic separation acting directly on the substance being separated
    • B03C1/28—Magnetic plugs and dipsticks
    • B03C1/288—Magnetic plugs and dipsticks disposed at the outer circumference of a recipient
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/42—Apparatus for the treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/50—Means for positioning or orientating the apparatus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
    • C12M33/06—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles for multiple inoculation or multiple collection of samples
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/06—Magnetic means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/04—Cell isolation or sorting
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/12—Purification
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
    • C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C2201/00—Details of magnetic or electrostatic separation
    • B03C2201/18—Magnetic separation whereby the particles are suspended in a liquid

Definitions

  • the present invention relates to a magnetic bead purification system for purification of substances such as proteins and nucleic acids.
  • Magnetic bead purification generally uses a purification system to separate magnetic beads by magnetic field to achieve separation and purification of substances such as cells, proteins or nucleic acids. Compared with the commonly used precipitation method, centrifugation method and column membrane method, the magnetic bead purification method has the characteristics of high extraction efficiency, fast separation speed and simple equipment.
  • the magnetic bead purification method based on magnetic adsorption can quickly and efficiently enrich the target protein in the sample by the combination of magnetic principle and incubation, which effectively avoids resin pretreatment and sample loading.
  • the limitations of the restrictions such as the constraints of packing and flow rate.
  • the purification method using magnetic separation is easier to automate, and can meet the high-throughput purification requirements of fast, automated, multi-channel simultaneous processing.
  • the only magnetic bead purification equipment currently on the market is the Kingfisher magnetic bead protein automatic purification instrument introduced by Thermo Scientific, which uses a magnetic bead method to insert a magnetic rod into the container for protein purification.
  • the maximum sample volume that can be purified is only 5ml.
  • the liquid transfer treatment requires manual participation, and the open-type work station can not control the pollution, which can not meet the customer's demand for automated processing of tens of milliliters and one hundred or two milliliters of sample processing. .
  • the magnetic bead purification system according to the present invention can easily realize large-volume sample processing.
  • a magnetic bead purification system comprising: an outer casing; a liquid path processing system disposed in the outer casing, the liquid path processing system being connectable with a reagent tank and a waste liquid tank; and the liquid path processing system a set of attached needles movable within the housing and coupled to the fluid handling system to receive reagents from the fluid handling system or to discharge waste fluid to the fluid handling system; Purifying a magnetic separation system comprising a magnetic element, the purified magnetic separation system being controllable to apply a lateral magnetic force or stop applying a magnetic force to a purification processing location in the outer casing by the magnetic element; and a purification station system capable of The purification processing location in the outer casing moves between a loading position outside the outer casing and the purification station system is adapted to load the container.
  • the magnetic element is a permanent magnet
  • the purified magnetic separation system controls the application of a magnetic force or stops the application of a magnetic force by moving the magnetic element close to and away from the purification processing position.
  • the purified magnetic separation system includes a drive system, a sliding system, and a transmission system to move the magnetic element closer to and away from the purification processing position.
  • the magnetic element is an electromagnet
  • the purified magnetic separation system controls the application of a magnetic force or stops the application of a magnetic force by controlling energization and de-energization of the electromagnet.
  • the purification station system includes a sliding system that is driven by the driving device to drive the purification station system from the purification processing position to the loading position.
  • the purification station system includes a magnetic block to move from the loading position to the purification processing position by the magnetic force of the magnetic element.
  • the purification magnetic separation applies a lateral magnetic force to attract the magnetic beads to the container wall of the container.
  • the purification station system comprises two purification station supports, and each purification station support comprises at least one matching hole of the purification container.
  • the loading needle set is a variable pitch loading needle set.
  • variable pitch sample pin set includes a fixed sample pin fixing block for mounting a sample needle and a mobile sample pin fixing block, the movable sample pin fixing block can be moved to adjust The spacing of the needles in the set of needles.
  • variable pitch syringe set comprises more than one mobile syringe pin block.
  • the set of loading needles comprises an elastic mechanism for moving the upper and lower movements of the loading needle.
  • the loading needle of the loading needle set includes a waste needle hole for discharging waste liquid and a sample needle hole for adding a reagent.
  • the loading pinhole is also used to spray the container wall and blow bubbles.
  • the magnetic bead purification system further comprises a mechanical arm disposed within the outer casing, the set of loading pins being mounted on the mechanical arm for movement.
  • the magnetic bead purification system comprises a cleaning module for cleaning the sample needle.
  • the cleaning module comprises a cleaning liquid supply hole, a liquid discharge hole, a cleaning tank and a cleaning liquid discharge tank matched with the sample needle.
  • the magnetic bead purification system further includes a control system that automatically controls operation of various components of the magnetic bead purification system in accordance with settings.
  • the magnetic bead purification system further comprises a pollution control system, the pollution control system comprising an air flow filtration system and an ultraviolet sterilization system.
  • the purification station system includes a fixture to load a 6-well container.
  • a magnetic bead purification system and components in accordance with a preferred embodiment of the present invention are exemplarily shown in the drawings.
  • the drawings are merely illustrative and the proportions are not necessarily consistent.
  • Figure 1 shows a magnetic bead purification system in accordance with one embodiment of the present invention.
  • FIG. 2 shows an internal structure diagram of a magnetic bead purification system in accordance with one embodiment of the present invention.
  • Figure 3 shows a schematic diagram of a fluid path processing system in accordance with one embodiment of the present invention.
  • FIGS. 4-1 through 4-3 illustrate a variable pitch loading pin set in accordance with an exemplary embodiment of the present invention.
  • Figure 5 illustrates a robotic arm in accordance with an exemplary embodiment of the present invention.
  • 6-1 and 6-2 illustrate a cleaning module in accordance with an exemplary embodiment of the present invention.
  • FIG. 7-1 through 7-4 illustrate a purified magnetic separation system and a purification station system in accordance with an exemplary embodiment of the present invention.
  • the invention discloses a semi-automated magnetic bead purification system.
  • the semi-automated magnetic bead purification system according to the present invention utilizes the magnetic bead method, combined with liquid transfer, blow mixing and incubation, magnetic bead adsorption and release, sample needle cleaning, air path filtration and ultraviolet sterilization, automatic extension and contraction of the working position. Back to the purification station and other functions, to achieve 12 samples of target protein incubation, washing, elution and other work.
  • the pollution control consists of a fully sealed safety cover, a safety door, a wind filter system, and an ultraviolet lamp.
  • the magnetic bead purification system may include the following modules: a casing 1, an opening and closing safety door 2, a control display screen 3, a wind path filtering system 4, a variable pitch loading needle set 5, The cleaning module 6, the purification magnetic separation system 7, the purification station system 8, the robot arm 9, the work substrate 10, the liquid processing system 11, and the control module 12.
  • the liquid path processing system may include a waste liquid tank 11-1, a two-way electromagnetic valve 11-2, a reagent barrel 11-3, and a six-turn.
  • the road valve 11-8 (converted from 7 liquid passages into one liquid passage), the liquid pipeline 11-9 and the waste liquid peristaltic pump connected to the sampling needle discharge passage are composed.
  • the drawings are merely exemplary schematic diagrams according to one embodiment of the present invention, and the liquid path treatment system may be adjusted according to a specific application, and the number and form of the solenoid valve, the liquid path valve, the waste liquid tank, and the like may be appropriately changed as needed. More specifically, the number of passages of the above liquid path valve can be appropriately changed depending on the specific application.
  • the liquid handling system can be used to add reagents, spray magnetic beads on the wall of the rinsing container and blow bubbles to mix the liquid, as well as clean the syringe, empty the waste container and clean the waste liquid in the module.
  • the liquid path treatment system includes at least one waste liquid peristaltic pump connected to the waste pinhole 5-6b, and the number of waste liquid peristaltic pumps connected to the waste pinhole 5-6b is the largest and the number of the load pins 5-6 same.
  • the liquid path treatment system includes at least one inlet peristaltic pump 11-5, and the number of the inlet peristaltic pumps 11-5 can be the same as that of the syringes 5-6.
  • the inlet peristaltic pump 11-5 is connected to the sample pinhole 5-6c.
  • the main part of the liquid handling system can be placed in the housing.
  • a waste liquid tank or the like may be disposed outside the outer casing, and a reagent source for supplying various reagents may be disposed outside the outer casing and connected to the liquid path processing system through a pipeline.
  • the two-way solenoid valve 11-2 is used to cut or connect the selected reagent liquid path.
  • Six-to-one liquid valve 11-4 which is composed of 6 liquid passages into one liquid passage, and with two-way solenoid valve 11-2, mainly used to select the required reagent or air, and the air is used for the needle Blow bubbles.
  • One-turn six-way valve 11-6 is divided into six liquid passages by one liquid passage, and is mainly used to distribute selected reagents or air to several syringes 5-6.
  • variable pitch loading needle set according to an exemplary embodiment of the present invention is illustrated.
  • the variable pitch type pin group is fixed to the Z axis of the robot arm 9, so that up and down and left and right movement can be realized.
  • the set of needles can also be a fixed pitch set of needles.
  • variable pitch loading needle set comprises at least one sampling needle 5-6, one elastic mechanism 5-9, a fixed loading needle fixing block 5-7, and a needle group bottom plate 5-8.
  • the variable pitch syringe set comprises a plurality of syringes 5-6.
  • variable pitch sample set When the variable pitch sample set includes two load pins 5-6 of variable pitch, it may also include a drive system 5-1, a mechanical limit 5-2, a control limit 5-4, and a sliding mechanism 5- 3 and the mobile syringe fixing block 5-5.
  • the loading needle set can flexibly realize the change of the spacing between the moving type needle fixing block 5-5 and the fixed type needle fixing block 5-7 by controlling the driving system and the guiding mechanism, and the adding needle can adapt to different hole spacing.
  • variable pitch syringe set includes more than 5 to 6 different needles of variable pitch
  • a plurality of mobile syringe fixing blocks 5-5 and corresponding drive, guide and control systems can be provided for flexibility
  • the distance between the movable sample pin fixing block 5-5 and the fixed sample pin fixing block 5-7 and the plurality of mobile sample pin fixing blocks 5-5 are changed, and the sample pin can be adapted to different hole spacings. .
  • the needles 5-6 may include a needle guiding mechanism 5-6a, a discharging needle hole 5-6b, and a loading needle hole 5-6c.
  • the elastic mechanism may include, for example, one spring and one guiding mechanism.
  • the drive system may for example comprise a hydraulic cylinder, a cylinder, or an electric motor.
  • the sliding mechanism can for example comprise a linear guide or a guide bushing.
  • the loading pins mounted on the same syringe retaining block can also be moved relative to each other to adjust the spacing.
  • the elastic mechanism is used for the upper and lower movement of the needle, and can fully touch the bottom of the container to completely process the liquid in the container without causing too much impact on the bottom of the container.
  • Discharged pinholes 5-6b are used to discharge waste liquid, and sample pinholes 5-6c are used to add reagents, spray the walls of the container and blow bubbles, and the reagents can rinse the magnetic beads on the walls of the container.
  • the sample needle is made of non-magnetic material and the surface is treated with hydrophobicity.
  • the waste pinhole 5-6b and the sample pinhole 5-6c of the sample needle 5-6 are respectively in communication with the corresponding passages of the liquid path treatment system.
  • the robot arm 9 includes at least a horizontally moving X-axis and a vertically moving Z-axis.
  • the Z axis of the robot arm 9 is used to secure the loading needle set.
  • the robot arm 9 can include two drive systems, two slide mechanisms, and two limit mechanisms for horizontal and vertical movement, respectively.
  • the vertically moving Z-axis drive system 9-1 is provided with a self-locking mechanism. The self-locking mechanism is used to ensure that the needle does not fall naturally under any circumstances.
  • the drive system may for example comprise a hydraulic cylinder, a cylinder or an electric motor.
  • the sliding mechanism can for example comprise a linear guide or a guide bushing.
  • the limit mechanism can include, for example, a limit switch and a mechanical limit block.
  • Figures 6-1 and 6-2 illustrate a cleaning module for cleaning the needles 5-6 in a fluid circuit system in accordance with a preferred embodiment of the present invention.
  • the cleaning module can for example be arranged on the working floor 10 of the magnetic bead purification system.
  • the cleaning module includes, for example, at least a cleaning tank 6-1, a cleaning liquid supply hole 6-2 (paired with the sample needle), a liquid discharge hole 6-4, and a cleaning liquid discharge tank 6-5, and can further Includes 1 external drain connector 6-3.
  • the number and arrangement of the cleaning dosing holes 6-2 correspond to the loading pins 5-6.
  • the needle When cleaning, the needle is inserted into the cleaning addition hole 6-2.
  • the liquid in the cleaning liquid filling hole flows out from the liquid discharging hole 6-4 at a lower speed than the filling liquid, so that the cleaning liquid is from the top of the cleaning liquid feeding hole.
  • the overflow liquid overflows from the cleaning liquid discharge tank 6-5 to the external liquid discharge interface 6-3, and the external liquid discharge interface 6-3 is connected with the waste liquid pump 11-7, so that the liquid in the cleaning module is discharged to the waste. Liquid pain, the purpose of cleaning the needle.
  • Figure 7-2 illustrates a purified magnetic separation system 7 in a magnetic bead purification system in accordance with a preferred embodiment of the present invention.
  • the purified magnetic separation system 7 may include a magnetic element 7-1 that generates a magnetic field, a fixed plate 7-2, a sliding mechanism 7-3, a drive system 7-4, a transmission system 7-5, and a limit.
  • the position mechanism 7-6 is composed.
  • the size and number of the magnetic elements 7-1 can be selected according to the needs of the application, and may be, for example, permanent magnets (magnets), electromagnets or the like.
  • three magnetic elements 7-1 are provided, which are respectively disposed at both sides and intermediate positions with respect to the moving direction of the purification station system.
  • the magnetic element 7-1 can be moved
  • the purification treatment position is here to apply a magnetic force.
  • the magnetic force applied by the magnetic element 7-1 to the purification processing position acts on the magnetic beads in the container, thereby adsorbing the magnetic beads (e.g., adsorbing the magnetic beads onto the container wall) for subsequent subsequent operations.
  • the arrangement of the magnetic elements 7-1 may be different, for example, may be set such that the width direction thereof is perpendicular to the moving direction of the purification station system 8, and when the purification station system 8 is moved to the purification processing position, the magnetic element 7 The -1 can be moved against the rear side of the purification station system 8. Such an arrangement is also allowed as long as the magnetic force is sufficient.
  • the magnetic element 7-1 When it is desired to stop the adsorption of the magnetic beads, the magnetic element 7-1 can be moved away from the purification processing position, thereby moving away from the magnetic beads to stop the application of the magnetic force. As long as the magnetic element 7-1 is far enough to stop the adsorption of the magnetic beads, it can be considered that the application of the magnetic force is stopped, although it is still possible that a weak magnetic force acts on the magnetic beads or acts on the purification processing position.
  • the purification treatment position refers to the area where the purification station system loading container is subjected to purification treatment in the outer casing, as opposed to the loading position of the loading container outside the outer casing.
  • the pure chemical system can be moved between the purification processing position in the housing and the loading position outside the housing.
  • the magnetic element 7-1 can be an electromagnet that can be positioned around the purification processing location to control the application and stop of magnetic forces by controlling its energization and de-energization.
  • the drive system may for example comprise a hydraulic cylinder, a cylinder, or an electric motor.
  • the sliding mechanism can for example comprise a linear guide or a guide bushing.
  • the limit mechanism can include, for example, a limit switch and a mechanical limit block.
  • the purification station system 8 includes a purification station holder fixing module 8-2 and a purification station holder 8-1.
  • the pure chemical position bracket 8-1 includes at least one purification station support, the upper side matching fixing hole 8-1a of one container, the lower side matching fixing hole 8-1b of one container, and the direction of the purification station holder.
  • the purification station holder 8-1 includes a plurality of purification station supports, and an upper side fitting fixing hole 8-1a of the corresponding number of containers and a lower side fitting fixing hole 8-1b of the container.
  • Each purification station can be placed in a container containing a mixture of the crude protein and magnetic beads after incubation.
  • the pure chemical position bracket fixing module 8-2 includes at least one purification station bracket fixing block 8-2a, one purification station bracket fixing block 8-2b, one purification station bracket fixing block 8-2c, one magnetic Suction block 8-2d (material is material that can be adsorbed by magnet, such as magnet, carbon steel, etc.), 1 sliding mechanism 8-2e, 1 limit block 8-2f, 1 movable magnetic limit block 8- 2g and 1 fixed magnetic limit block 8-2h.
  • Pure chemical position bracket fixing block 8-2a, purification station bracket fixing block 8-2b and purification station bracket fixing block 8-2c can cooperate to effectively fix the purification station bracket, and the direction fixing block of the purification station bracket 8-1c
  • the correct placement of the purification station bracket can be directed.
  • the number and position of the pure chemical support block 8-2a, the purification station support block 8-2b, and the purification station support block 8-2c can be determined according to the number of the purification station support 8-1 to be fixed.
  • each of the purification station holders 8-1 includes at least one matching hole of the purification container, and preferably may include six purification containers. Fit the hole.
  • the purification station system 8 can achieve a protein extraction station that extends from the purification station system to the outside of the instrument by purifying the motional thrust of the magnetic separation system 7 and the sliding mechanism 8-2e.
  • the pure chemical system 8 is pulled back from the outside of the instrument to the purification station by the suction of the magnetic element 7-1 in the magnetic block 8-2d and the magnetic separation system 7.
  • the suction of the movable magnetic limiting block 8-2g and the fixed magnetic limiting block 8-2h cooperates to return the purification station system to the purification position.
  • the drive unit and the guide rails can also be separately provided for the purification station system 8, and the push-out and pull-back movements can be performed by the control system.
  • the purification station system 8 does not require the provision of a purification station holder 8-1 that can be loaded directly onto the purification station holder fixing module 8-2 by means of a fixture.
  • the limit switch on the safety opening and closing door is used to control the opening and closing state of the door. If the safety opening and closing door is accidentally opened during the experiment, the experimental process is terminated. When ultraviolet sterilization is performed, if the opening and closing door is accidentally opened, the ultraviolet lamp is extinguished.
  • the purification process of the semi-automated magnetic bead purification system mainly includes experimental preparation, endotoxin control and purification procedures.
  • the reagents used in the following operations mainly include: reagent A: washing liquid for removing impurities in the liquid (components other than the target protein); reagent B: washing liquid, further removing impurities for removing the liquid, and Changing the liquid acid-base environment; reagent C: eluent for separating magnetic beads and target proteins; and reagent D: alkali solution for components such as bacteria in liquid lines and syringes in liquid systems.
  • the experimental preparation can mainly include:
  • endotoxin control can include:
  • the line is filled with D reagent, soaked, and the UV lamp is turned on at the same time.
  • the purification process can include:
  • the magnet is placed close to the centrifuge tube or the 6-well container to attract the magnetic beads.
  • the liquid is bubbled intermittently with 6 needles in the liquid of 6 centrifuge tubes or 6-well containers.
  • the magnet continues to adsorb the magnetic beads to promote the full adsorption of the magnetic beads.
  • the magnet is placed close to the centrifuge tube or the 6-well container to attract the magnetic beads.
  • the liquid is bubbled intermittently with 6 needles in the liquid of the six centrifuge tubes or the six-hole container.
  • the magnet continues to adsorb the magnetic beads to promote the full adsorption of the magnetic beads.
  • the magnet is placed close to the centrifuge tube or the 6-well container to attract the magnetic beads.
  • the liquid is bubbled intermittently with 6 needles in the liquid of the six centrifuge tubes or the six-hole container.
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Abstract

本发明提供一种磁珠纯化系统，其包括：外壳；设置在外壳中的液路处理系统，所述液路处理系统能够与试剂桶和废液桶连接；与所述液路处理系统连接的加样针组，所述加样针组能够在外壳内移动并且连接到所述液路处理系统，以从所述液路处理系统接收试剂或者向所述液路处理系统排出废液；纯化磁分离系统，其包括磁元件，所述纯化磁分离系统能够被控制以通过所述磁元件向外壳中的纯化处理位置处施加侧向的磁力或者停止施加磁力；和纯化工位系统，能够在外壳中的纯化处理位置与外壳外的装载位置之间移动，所述纯化工位系统适于装载容器。本发明的磁珠纯化系统能够容易实现大容量的样品处理。

Description

磁珠纯化系统 技术领域
本发明涉及一种用于蛋白质及核酸等物质的纯化的磁珠纯化系统。
背景技术
磁珠纯化法通常使用纯化系统通过磁场分离磁珠而达到分离和纯化细胞、蛋白或核酸等物质的目的。与常用的沉淀法、离心法、柱膜法相比，磁珠纯化法具有提取效率高、分离速度快、所需设备简单等特点。
与树脂纯化(Resin)相比，基于磁力吸附的磁珠纯化法依靠磁力原理和孵育的结合方式可以快速有效的富集样品中的目标蛋白，有效避免了树脂对样品预处理和对上样方式的限制等弊端，摆脱了装柱和流速等的限制。采用磁分离的纯化方法较易实现自动化，可满足快速、自动化、多通道同时处理的高通量纯化需求。
目前市面上仅有的磁珠纯化设备是Thermo Scientific公司推出的Kingfisher磁珠蛋白自动纯化仪，其利用的磁珠法采用磁棒插入到容器内部进行蛋白质的纯化。但其能纯化的最大样品体积只有5ml，液体转移处理需人工参与，并且采用敞口式的工作工位，不能控制污染，不能满足客户对自动化实现几十毫升和一两百毫升样本处理的需求。
因此，存在对磁珠纯化系统进行改进的需求。
发明内容
根据本发明的磁珠纯化系统能够容易实现大容量的样品处理。
根据本发明，公开了一种磁珠纯化系统，包括：外壳；设置在外壳中的液路处理系统，所述液路处理系统能够与试剂桶和废液桶连接；与所述液路处理系统连接的加样针组，所述加样针组能够在外壳内移动并且连接到所述液路处理系统，以从所述液路处理系统接收试剂或者向所述液路处理系统排出废液；纯化磁分离系统，包括磁元件，所述纯化磁分离系统能够被控制以通过所述磁元件向外壳中的纯化处理位置处施加侧向的磁力或者停止施加磁力；和纯化工位系统，能够在外壳中的纯化处理位置与外壳外的装载位置之间移动，所述纯化工位系统适于装载容器。
优选的，所述磁元件为永磁体，所述纯化磁分离系统通过移动所述磁元件接近和远离所述纯化处理位置来控制施加磁力或者停止施加磁力。
优选的，所述纯化磁分离系统包括驱动系统、滑动系统和传动系统，以移动所述磁元件接近和远离所述纯化处理位置。
优选的，所述磁元件为电磁铁，所述纯化磁分离系统通过控制所述电磁铁的通电和断电来控制施加磁力或者停止施加磁力。
优选的，所述纯化工位系统包括滑动系统，通过驱动装置以驱动所述纯化工位系统从所述纯化处理位置移动到所述的装载位置。
优选的，所述纯化工位系统包括磁吸块，以通过所述磁元件的磁力从所述的装载位置移动到所述纯化处理位置。
优选的，当装有孵育后粗蛋白和磁珠的混合液的容器安装在所述纯化工位系统上，并且所述纯化工位系统移动到所述纯化处理位置中时，所述纯化磁分离系统施加侧向的磁力以将磁珠吸附在所述容器的容器壁上。
优选的，所述纯化工位系统包括2个纯化工位支架，每个纯化工位支架分别包括至少1个纯化容器的适配孔。
优选的，所述加样针组为可变间距加样针组。
优选的，所述可变间距加样针组包括用于安装加样针的固定式加样针固定块和移动式加样针固定块，所述移动式加样针固定块能够被移动以调节所述加样针组内加样针的间距。
优选的，所述可变间距加样针组包括多于1个移动式加样针固定块。
优选的，所述加样针组包括用于加样针的上下微量移动的弹性机构。
优选的，所述加样针组的加样针包括用于排废液的排废针孔和用于添加试剂的加样针孔。
优选的，所述加样针孔还用于喷洒冲洗容器壁和吹打气泡。
优选的，所述磁珠纯化系统还包括在外壳内设置的机械臂，所述加样针组安装在所述机械臂上以实现移动。
优选的，所述磁珠纯化系统包括清洗加样针的清洗模块。
优选的，所述清洗模块包括与加样针配对的清洗加液孔、排液孔、清洗槽和清洗排液槽。
优选的，所述磁珠纯化系统还包括控制系统，所述控制系统根据设置自动控制所述磁珠纯化系统的各个部件的操作。
优选的，所述磁珠纯化系统还包括污染控制系统，所述污染控制系统包括风路过滤系统和紫外杀菌系统。
优选的，所述纯化工位系统包括固定装置以装载6孔容器。
附图说明
附图中示例性地示出了根据本发明的优选实施例的磁珠纯化系统及部件。各附图只是示例性说明，其比例不必要一致。
图1示出了根据本发明一个实施例的磁珠纯化系统。
图2示出了根据本发明一个实施例的磁珠纯化系统的内部结构图。
图3示出了根据本发明一个实施例的液路处理系统的原理图。
图4-1至图4-3示出了根据本发明的示例性实施例的可变间距加样针组。
图5示出了根据本发明的示例性实施例的机械臂。
图6-1和图6-2示出了根据本发明示例性实施例的清洗模块。
图7-1至图7-4示出了根据本发明示例性实施例的纯化磁分离系统和纯化工位系统。
具体实施方式
为了更清楚地说明本发明实施例的技术方案，下面将对照附图说明本发明的具体实施方式。显而易见地，下面描述中的附图仅仅是本发明的一些示例性实施例。
本发明公开了一种半自动化的磁珠纯化系统。根据本发明的半自动化的磁珠纯化系统利用磁珠法，配合液体转移、吹打混匀孵育、磁珠吸附及释放、加样针清洗、风路过滤及紫外杀菌、工作位自动伸出和缩回到纯化工位等功能，实现12个样本的目标蛋白孵育、洗杂、洗脱等工作。污染控制由全密封安全罩、安全门、风路过滤系统、紫外灯等组成。
如图1和图2所示，根据本发明的磁珠纯化系统可以包括如下模块：外壳1、开合安全门2、控制显示屏3、风路过滤系统4、可变间距加样针组5、清洗模块6、纯化磁分离系统7、纯化工位系统8、机械臂9、工作底板10、液体处理系统11和控制模块12。
在图3中，示出了根据本发明的液路处理系统的原理图，此液路处理系统可以包括废液桶11-1、二通电磁阀11-2、试剂桶11-3、六转一液路阀11-4、进液蠕动泵11-5、一转六液路阀11-6、加样针5-6、排废液蠕动泵11-7、清洗模块6、七转一液路阀11-8(由7个液路通道汇成1个液路通道)、液路管11-9和加样针排液通道连接的排废液蠕动泵组成。附图只是根据本发明一个实施例的示例性原理图，可以根据具体应用对液路处理系统进行调整，例如电磁阀、 液路阀、废液桶等的数量和形式可以根据需要进行适当改变。更具体地，上述液路阀的通路数量可以根据具体应用进行适当改变。
液路处理系统可以用于添加试剂、喷洒冲洗容器管壁上的磁珠和吹打气泡用于混匀液体，以及清洗加样针、排空容器内和清洗模块内的废液等。
液路处理系统至少包括1个与排废针孔5-6b连接的排废液蠕动泵，排废针孔5-6b连接的排废液蠕动泵的数量最多与加样针5-6的数量一样。
液路处理系统至少包括1个进液蠕动泵11-5，进液蠕动泵11-5的数量可与加样针5-6一样。进液蠕动泵11-5与加样针孔5-6c连接。
液路处理系统的主要部分可以设置在外壳中。优选的，废液桶等也可以设置在外壳之外，供应各种试剂的试剂源也可以设置在外壳之外，通过管路与液路处理系统相连。
二通电磁阀11-2，用于切断或者连通所选择的试剂液路通道。
六转一液路阀11-4，由6个液路通道汇成1个液路通道，配合二通电磁阀11-2，主要用于选择所需要的试剂或者空气，空气用于加样针吹打气泡。
一转六液路阀11-6，由1个液路通道分成6个液路通道，主要用于将选择的试剂或空气分配到若干个加样针5-6。
在图4-1至图4-3中，示出了根据本发明的示例性实施例的可变间距加样针组。如图4-1至图4-3所示，可变间距加样针组固定在机械臂9的Z轴上，从而可以实现上下和左右移动。在某些实施例中，加样针组也可以是固定间距的加样针组。
可变间距加样针组包括至少1个加样针5-6、1个弹性机构5-9、固定式加样针固定块5-7、针组底板5-8组成。优选的，可变间距加样针组包括多个加样针5-6。
当可变间距加样针组包括可变间距的2个加样针5-6时，还可以包括驱动系统5-1、机械限位5-2、控制限位5-4、滑动机构5-3和移动式加样针固定块5-5。加样针组通过控制驱动系统、导向机构可以灵活实现移动式加样针固定块5-5与固定式加样针固定块5-7间的间距变化，加样针可以适应不同的孔间距。
当可变间距加样针组包括可变间距的更多个加样针5-6时，可以设置多个移动式加样针固定块5-5和相应的驱动、导向及控制系统，从而灵活实现移动式加样针固定块5-5与固定式加样针固定块5-7间以及多个移动式加样针固定块5-5间的间距变化，加样针可以适应不同的孔间距。
加样针5-6可以包括加样针导向机构5-6a、排废针孔5-6b和加样针孔5-6c。
弹性机构例如可以包括1个弹簧和1个导向机构。
驱动系统例如可以包括液压缸、气缸、或电动马达。
滑动机构例如可以包括直线导轨或导轴导套。
在某些实施例中，同一加样针固定块上安装的加样针也可以相对彼此移动以调节间距。
弹性机构用于加样针的上下微量移动，能够充分触到容器底部，完全处理容器内的液体，同时不会对容器底部造成太大冲击。
排废针孔5-6b用于排废液，加样针孔5-6c用于添加试剂、喷洒冲洗容器管壁和吹打气泡，试剂可以冲洗容器壁上的磁珠。加样针选用无磁材料，表面进行疏水处理。加样针5-6的排废针孔5-6b和加样针孔5-6c分别与液路处理系统的相应通道连通。
如图5所示，机械臂9至少包括水平移动的X轴和垂直移动的Z轴。机械臂9的Z轴用于固定加样针组。机械臂9可包括2个驱动系统、2个滑动机构和2个限位机构，分别用于水平方向和垂直方向的移动。其中，垂直移动的Z轴的驱动系统9-1带有自锁机构。自锁机构用于保证加样针在任何情况下都不会自然下落。
驱动系统例如可以包括液压缸、气缸或电动马达。
滑动机构例如可以包括直线导轨或导轴导套。
限位机构例如可以包括限位开关和机械限位块。
图6-1和图6-2示出了根据本发明优选实施例的清洗模块，用于清洗液路系统中的加样针5-6。该清洗模块例如可以布置在磁珠纯化系统的工作底板10上。清洗模块例如至少包括清洗槽6-1、1个清洗加液孔6-2(与加样针配对)、1个排液孔6-4、1个清洗排液槽6-5，并可以进一步包括1个外置排液接口6-3。优选的，清洗加液孔6-2的数量和布置对应于加样针5-6。
清洗时，加样针插入清洗加液孔6-2。由于加样针加入清洗液到清洗加液孔6-2时，清洗加液孔内的液体从排液孔6-4流出的速度低于加液的速度，使清洗液从清洗加液孔顶部溢出，溢出液体从清洗排液槽6-5流到外置排液接口6-3，外置排液接口6-3与排废液泵11-7连接，实现清洗模块内的液体排到废液痛，达到清洗加样针的目的。
图7-2示出了根据本发明优选实施例的磁珠纯化系统中的纯化磁分离系统7。如图7-2所示，纯化磁分离系统7可以包括产生磁场的磁元件7-1、固定板7- 2、滑动机构7-3、驱动系统7-4、传动系统7-5、和限位机构7-6组成。磁元件7-1的大小和数量可以根据应用的需求进行选择，其例如可以是永磁体(磁铁)、电磁铁等。
根据图7-1至图7-4中所示的实施例，设置有3个磁元件7-1，相对于纯化工位系统的移动方向分别设置在两侧和中间位置。当将容纳有孵育后的粗蛋白和磁珠的混合液的容器装载到纯化工位系统8并将纯化工位系统8移动到外壳中的纯化处理位置处时，可以将磁元件7-1移动到此纯化处理位置处以施加磁力。磁元件7-1向纯化处理位置处施加的磁力作用于容器中的磁珠上，从而吸附磁珠(例如将磁珠吸附到容器壁上)，以进行相应的后续操作。
在其他实施例中，磁元件7-1的布置可以不同，例如可以设置为其宽度方向垂直于纯化工位系统8的移动方向，当纯化工位系统8移动到纯化处理位置时，磁元件7-1可以被移动贴靠纯化工位系统8的后侧。只要磁力足够，这样的布置也是允许的。
当需要停止吸附磁珠时，可以将磁元件7-1移动远离纯化处理位置，从而远离磁珠以停止施加磁力。只要磁元件7-1足够远以停止吸附磁珠，即可视为停止施加磁力，尽管仍然可能有微弱磁力作用在磁珠上或者作用于纯化处理位置处。
纯化处理位置是指纯化工位系统装载容器在外壳中进行纯化处理的区域，与其在外壳之外装载容器的装载位置相对。纯化工位系统可在外壳中的纯化处理位置和外壳之外的装载位置之间移动。
在其他实施例中，磁元件7-1可以是电磁铁，其位置可以固定在纯化处理位置周围，通过控制其通电和断电来控制磁力的施加和停止。
驱动系统例如可以包括液压缸、气缸、或电动马达。
滑动机构例如可以包括直线导轨或导轴导套。
限位机构例如可以包括限位开关和机械限位块。
如图7-1至7-4所示，纯化工位系统8包括纯化工位支架固定模块8-2和纯化工位支架8-1。纯化工位支架8-1至少包括1个纯化工位支撑、1个容器的上侧适配固定孔8-1a、1个容器的下侧适配固定孔8-1b和纯化工位支架的方向固定块8-1c。优选的，纯化工位支架8-1包括多个纯化工位支撑，以及相应数量的容器的上侧适配固定孔8-1a和容器的下侧适配固定孔8-1b。每个纯化工位可以放入装有孵育后的粗蛋白和磁珠混合液的容器。
纯化工位支架固定模块8-2至少包括1个纯化工位支架固定块8-2a、1个纯化工位支架固定块8-2b、1个纯化工位支架固定块8-2c、1个磁吸块8-2d(材 料为可以被磁铁吸附的材料，如磁铁、碳钢等)、1个滑动机构8-2e、1个限位块8-2f、1个移动式磁性限位块8-2g和1个固定磁性限位块8-2h。
纯化工位支架固定块8-2a、纯化工位支架固定块8-2b和纯化工位支架固定块8-2c可以协作以有效固定纯化工位支架，纯化工位支架的方向固定块8-1c可以指引正确放置纯化工位支架。纯化工位支架固定块8-2a、纯化工位支架固定块8-2b和纯化工位支架固定块8-2c的数量和位置可以根据需要固定的纯化工位支架8-1的数量确定。
在图示的实施例中，固定有两个纯化工位支架8-1，每个纯化工位支架8-1分别包括至少1个纯化容器的适配孔，优选的可以包括6个纯化容器的适配孔。
在图示的实施例中，纯化工位系统8可以通过纯化磁分离系统7的运动推力和滑动机构8-2e，实现纯化工位系统伸出到仪器外侧的蛋白手工提取工位。纯化工位系统8通过磁吸块8-2d与磁分离系统7中的磁性元素7-1的吸力，从仪器外侧拉回到纯化工位。拉回运动的过程中，移动式磁性限位块8-2g和固定磁性限位块8-2h的吸力配合，实现纯化工位系统回到纯化位置。当然，也可以为纯化工位系统8单独设置驱动装置和导轨，并通过控制系统进行推出和拉回运动。
根据其他实施例，纯化工位系统8不需要设置纯化工位支架8-1，其可以在纯化工位支架固定模块8-2上直接通过固定装置装载6孔容器。
安全开合门上有限位开关，用于控制门的开合状态，如果在实验过程中意外打开安全开合门，则实验流程终止。当进行紫外杀菌时，如果意外打开开合门，则紫外灯熄灭。
以下根据图示的实施例，简要说明根据本发明的磁珠纯化系统的实验流程或操作方法。
根据本发明的半自动化的磁珠纯化系统的纯化流程主要包括实验准备、内毒素控制和纯化流程。以下操作中使用的试剂主要包括：试剂A：洗杂液，用于去除液体中的杂质(除目标蛋白之外的成分)；试剂B：清洗液，进一步去除用于去除液体中的杂质，和改变液体酸碱环境；试剂C：洗脱液，用于分离磁珠和目标蛋白；以及试剂D：碱溶液，用于液体系统中液体管路和加样针中的细菌等成分。
第一，实验准备主要可以包括：
1.系统自检和复位。
2.排空液路中所有管路(包括试剂桶到二通电磁阀间的管路)。
第二，内毒素控制可以包括：
1.管路充满D试剂，浸泡，并且同时打开紫外灯。
2.用D试剂清洗加样针，并排空。
3.用B试剂清洗管路和加样针，并排空。
4.人工将装有孵育后的粗蛋白和磁珠混合液的12个离心管，或者1个或2个6孔容器放入纯化工位。
第三，纯化流程可以包括：
1.磁铁靠近离心管或者6孔容器，吸附磁珠。
2.在磁铁吸附的状态下，用6针向前6个离心管或者6孔容器的液体中，间断性吹气鼓泡。
3.磁铁继续吸附磁珠，促进磁珠充分吸附。
4.用6针进入前6个离心管或者6孔容器吸取上清至废液桶。
5.用A试剂清洗管路和针头。
6.重复1次同2-4的步骤，取完后6个离心管或者6孔容器中的上清，吸液期间针能间断下降。
7.用A试剂清洗管路和针头。
8.磁铁离开。
9.用6针对12个离心管或者6孔容器加A试剂第1次，分2次加完，加样针悬空加液，确保把磁珠完全冲洗下来。
10.用6针向前6个离心管或者6孔容器的液体中，间断性吹气鼓泡，进行充分混匀磁珠。
11.用A试剂清洗管路和针头。
12.用6针向后6个离心管或者6孔容器的液体中，间断性吹气鼓泡，进行充分混匀磁珠，静置。
13.磁铁靠近离心管或者6孔容器，吸附磁珠。
14.在磁铁吸附的状态下，用6针向后6个离心管或者6孔容器的液体中，间断性吹气鼓泡。
15.磁铁继续吸附磁珠，促进磁珠充分吸附。
16.用6针进入后6个离心管或者6孔容器吸取上清至废液桶。
17.用A试剂清洗管路和针头。
18.重复1次同14-16的步骤，取完前6个离心管或者6孔容器中的上清，吸液期间针能间断下降。
19.重复步骤5-16，反复3次。
20.用B试剂清洗管路和针头。
21.磁铁离开。
22.用6针对12个离心管或者6孔容器加B试剂，分2次加完，加样针悬空加液，确保把磁珠完全冲洗下来。
23.用6针向前6个离心管或者6孔容器的液体中，间断性吹气鼓泡，进行充分混匀磁珠，静置。
24.用B试剂清洗管路和针头。
25.用6针向后6个离心管或者6孔容器的液体中，间断性吹气鼓泡，进行充分混匀磁珠，静置。
26.磁铁靠近离心管或者6孔容器，吸附磁珠。
27.在磁铁吸附的状态下，用6针向后6个离心管或者6孔容器的液体中，间断性吹气鼓泡。
28.磁铁继续吸附磁珠，促进磁珠充分吸附。
29.用6针进入后6个离心管或者6孔容器吸取上清至废液桶。
30.用B试剂清洗管路和针头。
31.重复1次同27-29的步骤，取完前6个离心管或者6孔容器中的上清，吸液期间针能间断下降。
32.磁铁离开。
33.用C试剂清洗管路和针头。
34.用6针对前6个离心管或者6孔容器加C试剂第一次，加样针悬空加液，确保把磁珠完全冲洗下来。
35.加完C试剂后，立即用6针向前6个离心管或者6孔容器的液体中，间断性吹气鼓泡。
36.用C试剂清洗管路和针头。
37.用6针对后6个离心管或者6孔容器加C试剂第一次，加样针悬空加液，确保把磁珠完全冲洗下来。
38.加完C试剂后，立即用6针向前6个离心管或者6孔容器的液体中，间断性吹气鼓泡，之后静置。
39.磁铁靠近离心管或者6孔容器，吸附磁珠。
40.打开安全门，等待操作者手工转移蛋白。
41.完成手工转移蛋白后，关上安全门。
42.磁铁离开。
43.用C试剂清洗管路和针头。
44.用6针对前6个离心管或者6孔容器加C试剂第二次，加样针悬空加液，确保把磁珠完全冲洗下来。
45.加完C试剂后，立即用6针向前6个离心管或者6孔容器的液体中，间断性吹气鼓泡，之后静置。
46.用C试剂清洗管路和针头。
47.用6针对后6个离心管或者6孔容器加C试剂第二次，加样针悬空加液，确保把磁珠完全冲洗下来。
48.加完C试剂后，立即用6针向前6个离心管或者6孔容器的液体中，间断性吹气鼓泡，之后静置。
49.磁铁靠近离心管或者6孔容器，吸附磁珠。
50.打开安全门，等待操作者手工转移蛋白。
51.完成手工转移蛋白后，关上安全门。
52.磁铁离开。
53.用C试剂清洗管路和针头。
应当说明的是，上述实施例均可根据需要自由组合。以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干变化和改进，这些变化和改进也应视为落入本发明的保护范围。

Claims (20)

  1. 一种磁珠纯化系统，其特征在于包括：
    外壳；
    设置在外壳中的液路处理系统，所述液路处理系统能够与试剂桶和废液桶连接；
    与所述液路处理系统连接的加样针组，所述加样针组能够在外壳内移动并且连接到所述液路处理系统，以从所述液路处理系统接收试剂或者向所述液路处理系统排出废液；
    纯化磁分离系统，包括磁元件，所述纯化磁分离系统能够被控制以通过所述磁元件向外壳中的纯化处理位置处施加侧向的磁力或者停止施加磁力；和
    纯化工位系统，能够在外壳中的纯化处理位置与外壳外的装载位置之间移动，所述纯化工位系统适于装载容器。
  2. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述磁元件为永磁体，所述纯化磁分离系统通过移动所述磁元件接近和远离所述纯化处理位置来控制施加磁力或者停止施加磁力。
  3. 根据权利要求2所述的磁珠纯化系统，其特征在于，所述纯化磁分离系统包括驱动系统、滑动系统和传动系统，以移动所述磁元件接近和远离所述纯化处理位置。
  4. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述磁元件为电磁铁，所述纯化磁分离系统通过控制所述电磁铁的通电和断电来控制施加磁力或者停止施加磁力。
  5. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述纯化工位系统包括滑动系统，通过驱动装置以驱动所述纯化工位系统从所述纯化处理位置移动到所述的装载位置。
  6. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述纯化工位系统包括磁吸块，以通过所述磁元件的磁力从所述的装载位置移动到所述纯化处理位置。
  7. 根据权利要求1-6中任一项所述的磁珠纯化系统，其特征在于，当装有孵育后粗蛋白和磁珠的混合液的容器安装在所述纯化工位系统上，并且所述纯化工位系统移动到所述纯化处理位置中时，所述纯化磁分离系统施加侧向的磁力以将磁珠吸附在所述容器的容器壁上。
  8. 根据权利要求1、5和6中任一项所述的磁珠纯化系统，其特征在于，所述纯化工位系统包括2个纯化工位支架，每个纯化工位支架分别包括至少1个纯化容器的适配孔。
  9. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述加样针组为可变间距加样针组。
  10. 根据权利要求9所述的磁珠纯化系统，其特征在于，所述可变间距加样针组包括用于安装加样针的固定式加样针固定块和移动式加样针固定块，所述移动式加样针固定块能够被移动以调节所述加样针组内加样针的间距。
  11. 根据权利要求10所述的磁珠纯化系统，其特征在于，所述可变间距加样针组包括多于1个移动式加样针固定块。
  12. 根据权利要求1和9-11中任一项所述的磁珠纯化系统，其特征在于，所述加样针组包括用于加样针的上下微量移动的弹性机构。
  13. 根据权利要求1和9-11中任一项所述的磁珠纯化系统，其特征在于，所述加样针组的加样针包括用于排废液的排废针孔和用于添加试剂的加样针孔。
  14. 根据权利要求13所述的磁珠纯化系统，其特征在于，所述加样针孔还用于喷洒冲洗容器壁和吹打气泡。
  15. 根据权利要求1和9-11中任一项所述的磁珠纯化系统，其特征在于，所述磁珠纯化系统还包括在外壳内设置的机械臂，所述加样针组安装在所述机械臂上以实现移动。
  16. 根据权利要求1和9-11中任一项所述的磁珠纯化系统，其特征在于，所述磁珠纯化系统包括清洗加样针的清洗模块。
  17. 根据权利要求16所述的磁珠纯化系统，其特征在于，所述清洗模块包括与加样针配对的清洗加液孔、排液孔、清洗槽和清洗排液槽。
  18. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述磁珠纯化系统还包括控制系统，所述控制系统根据设置自动控制所述磁珠纯化系统的各个部件的操作。
  19. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述磁珠纯化系统还包括污染控制系统，所述污染控制系统包括风路过滤系统和紫外杀菌系统。
  20. 根据权利要求1所述的磁珠纯化系统，其特征在于，所述纯化工位系统包括固定装置以装载6孔容器。
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