SU1124976A1 - Method of obtaining sorbent - Google Patents

Description

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The invention relates to a vaccine serum case and can be used in the technology for obtaining erythrocyte diagnostics of medical and medical. veterinary prescribing.

Erythrocyte selection methods for obtaining erythrocyte diagnosticum are not known in the patent and scientific literature.

The aim of the invention is to create a method for selecting red blood cells to obtain an erythrocyte diagnosticum.

The goal is achieved by the fact that, according to the method of selecting red blood cells to obtain an erythrocyte diagnosticum, which consists in that red blood cells are separated from donor blood, fixed, then the charge of fixed red blood cells is determined and erythrocytes with a charge of 4.12-10 are used as the starting material - 3, Cl.

The method is carried out as follows. After obtaining whole blood (e.g. lamb), erythrocytes are extracted from it, then they are fixed with formalin and subjected to analytical or preparative cell electrophoresis, charge is determined, and further erythrocytes with charge of antigens are used as particle carriers of antigens. 4.1210-3, Cl.

In the technical implementation of the proposed method for selection of erythrocytes, the limiting value of the charge value is preliminarily established, with a decrease in which for fixed erythrocytes, erythrocytes are screened. Boundary values can be determined by a single determination of the charge of fixed erythrocytes; with parallel monitoring of diagnostic kits made on these erythrocytes. According to the Production Regulations, the quality control of diagnosticum is carried out in the reaction of RIGA with the hyperimmune sera of animals and the sera of healthy donors, i.e. by graduation, as illustrated in the example. Moreover, according to the technical regulations of the Production Regulations, the diagnosticum is considered specific if its titer with serum of healthy donors in the RIGA reaction is less than 1/100.

Example. Selection of fixed erythrocytes 1 ml of formalized sheep erythrocytes contained in physiological saline is precipitated by centrifugation at 350 for 5 min, followed by washing twice and resuspension in a measuring medium with the parameters: ionic strength (u 0.02; rI 7.3;; erythrocyte concentration; erythrocyte concentration; in the sample, it is diluted with a measuring medium to 5-10 5-10 cells per 1 ml. The surface charge of the erythrocyte is calculated using the formula g 3-lG 4. (i5 CG5E and US C, where the measured value is U electrophoretic mobility st (efp), μm C-viscosity (for solutions used is 10 n), the value is the inverse of the thickness of the electrical double layer, which for aqueous solutions of monovalent salts, (concentration. cm) is equal to e-cm / 3. cm; S is the surface area of the errocyte, the average value of which is 1, 65-ICT cm.

AFP is measured by the method of microelectrophoresis on a Parmoquant-2 instrument (Carl Zeis, Jena) in an automatic mode. The instrument used provides the average values of the ESP and statistical data processing. Comparative data of the obtained EFP of erythrocytes in comparison with the determination of the specificity of drugs (made on these erythrocytes) by the reaction of RIGA are given in table.

Eight batches of formalized erythrocytes (column 1) compared with RIGA titers of diagnostics made on these erythrocytes (columns 2 and 3), the first four batches of both liquid and dry preparations have titers greater than 1/100, those. according to the technical conditions of the Production Regulations, they must be rejected. All these batches of red blood cells EFR less than 2 microns C In see Preparations of the parties, 5-8 have a titer less than 1/100, i.e. correspond to the specifications. The ESP of these batches is more than 2 μm V cm. Moreover, the preparation, the erythrocytes of which have an ESP of 1.93 μm cm, has a titer of 1/150 and 1/300, i.e. are bad and the preparation, whose EFP of 2.02 µm cm has a titer of 1/50, i.e. are good. Thus, the lower bound of the values of the ESP, at which it is necessary to reject the erythrocytes, is 2 µm C – B cm or 3.3–10 Kl.

The upper limit of the EFP values of fixed erythrocytes is determined by the nature of the surface of the native erythrocytes of the ram. This value was found to be 2.5 µm Svcm or

4-1210 Cl.

Visual control spontaneous

agglutination does not allow detection of bad red blood cells and diagnosticums are rejected after their manufacture, and the use of the proposed method prevents the release of non-specific diagnosticum. In addition, the proposed method allows the rejection of red blood cells quickly, immediately after their fixation, i.e. there is no need for long-term production of diagnostic kits on these erythrocytes and staging RIGA with donor sera.

Claims (1)

57) METHOD FOR SELECTING RYTHROCYTES TO OBTAIN. Erythrocyte diagnostics, consisting in the fact that red blood cells are isolated from the donor; blood, they are fixed, then the charge of fixed red blood cells is determined and erythrocytes with a charge of 4.12 ΊΟ ' ¹ 3.3Ί0 ^η Cl are used as feedstock. The invention relates to a vaccine-serum case and can be used in the technology of obtaining red blood cell diagnosticum medical. veterinary appointment. In the patent and scientific literature, there are no known methods for the selection of red blood cells to obtain a red blood cell diagnosticum. The aim of the invention is to provide a method for the selection of red blood cells to obtain a red blood cell diagnosticum. This goal is achieved by the fact that according to the method of selecting red blood cells to obtain an erythrocyte diagnosticum, which consists in the fact that red blood cells are extracted from donated blood, they are fixed, then the charge of fixed red blood cells is determined and red blood cells with a charge of 4.12 · 10 ** ² - are used as feedstock 3,3-NU ' ¹ Cl. The method is as follows. After obtaining whole blood (for example, lamb), red blood cells are extracted from it, then they are fixed with formalin and subjected to analytical or preparative cell electrophoresis, the charge is determined, and erythrocytes with a charge of 4.12 are used as particles-carriers of antigens in the further production of diagnosticum from the whole population · 10 “ ^{! Ζ} - 3.3 · 10 ~ ' ² ' Cl. In the technical implementation of the proposed method for the selection of red blood cells, a boundary value of the charge value is preliminarily set, upon reduction of which red blood cells are discarded in fixed red blood cells. Boundary values can be determined by a single determination of the charge of fixed red blood cells: with parallel monitoring of diagnostics made on these red blood cells. According to the Production Regulations, the quality control of the diagnosticum is carried out in the reaction of ΡΗΓΑ with hyperimmune animal sera and healthy donor sera, i.e. by graduation, as illustrated in the example. Moreover, according to the technical conditions of the Production Regulations, a diagnosticum is considered specific if its titer with healthy donor serum in the ΡΗΓΑ reaction is less than 1/100. Example. The sampling of fixed red blood cells 1 ml of formalized sheep erythrocytes contained in physiological saline is precipitated by centrifugation at £ = · 350 for 5 min, followed by two times washing and resuspension in a measuring medium with the following parameters: ionic strength p = 0.02; pH 7.3; t = 25 ° C. The concentration of red blood cells in the sample was adjusted by diluting it with a measuring medium to 5 · 1Ό ⁶ 5 · 10 ² cells per 1 ml. The surface charge of the red blood cell is calculated by the formula g = 3 · 10 ' ^{T |} 7 “U5 CG 5Е = 10 '” J those U5 C, where the measured quantity is U electrophoretic mobility (EF), μm C * B *'cm; - viscosity (for used solutions is equal to 10 ^_g p); % is the reciprocal of the thickness of the double electric layer, which for aqueous solutions of monovalent salts (concentration, cm) is equal to eGeSm / W · 10® cm ' ¹ 10 ^{+ g} cm ~ *; S is the surface area of the red blood cell, the average value of which is 1.65 · KG ^ cm ² . EFP is measured by the method of microelectrophoresis on an instrument ¹¹ Parmokvant-2 ' ¹ (' Karl Zeise ¹ ', Jena) in automatic mode. The instrument used provides the average values of the EFP and statistical data processing. Comparative data of the obtained EPC of erythrocytes in comparison with the determination of the specificity of the preparations (made on these erythrocytes) by the reaction ΡΗΓΑ are given in table. According to the data of the EFP control of eight batches of formalized erythrocytes (column 1) in comparison with the ΑΓΑ titers of diagnosticums made on these erythrocytes (columns. 2 and 3), the first four batches of both liquid and dry preparations have titers greater than 1/100 , i.e. according to the technical conditions of the production regulations they must be rejected. All of these batches of erythrocytes have EF (1. Less than 2 μm C V cm. The preparations of batches 5-8 have a titer less than 1/100, i.e. they correspond to TU. the preparation, the red blood cells of which have an EFP of 1.93 μm C ' ¹ B' ¹ cm, have a titer of 1/150 and 1/300, i.e. they are bad, and the drug, the EFP of which 2.02 μm C ' ¹ cm have a titer 1/50, i.e. are good. Thus, the lower limit of the values of the electron phase transition at which it is necessary to carry out the selection. erythrocyte forging, equal to 2 μm C ^m V ”cm or 3.3 - 10 '^ C. The upper limit of the EFP values of fixed erythrocytes is determined by the nature of the surface of native sheep erythrocytes. This value turned out to be equal to 2.5 μm s'v'cm or 4 · 12'10 ' ² C. Visual control of spontaneous agglutination does not allow to identify bad red blood cells and diagnostics are rejected after their manufacture, as well. the use of the proposed method prevents the release of a non-specific diagnosticum. In addition, the proposed method allows the rejection of red blood cells quickly, immediately after their fixation, i.e. Otpa gives the need for long-term production of diagnosticums on these red blood cells and the setting of ΡΗΓΑ with donor sera. EFP formalin- Title of diagnosticum in ΡΗΓΑ with donor pp erythrocytes serum cit, μm C * ’B“ ’cm liquid preparation dry preparation 11.41 21.47 31.59 41.93 1/1200 1/1200 1/300 1/300 1/1200 1/1200 1/600 1/150 52.02 62.13 72.23 82.31 1/50 1/50 1/50 1/50 0