RU2429015C2 - Method of preparing product containing immunoglobulins g, a, m (versions) - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: what is offered is a method for producing a complex of immunoglobulins G, A, M. A starting material is deposited B(III), A(II+III) Cohn fractions or their mixtures in any proportion which are homogenised in 0.3-0.7 % sodium chloride to produce a suspension which is processed by 1-3 % chloroform at the pH value 4.9-5.6, mixed, kept for 30-60 min, centrifuged, and an end product is recovered with removing excess liquid. Another version of the method provides homogenising the starting material in 0.3-0.7 % sodium chloride to produce a suspension which is processed by 1-3 % chloroform at pH 4.9-5.6, mixed; the suspension is kept for 30-60 min, centrifuged, and then spend a stage of additional deposition of ballast fractions in a solution of ionic strength less than 0.05 at pH 4.5 to 5.6 for 12-24 hours followed by removal by additional centrifugation or filtration, and then an end product is recovered by ultrafiltration.

EFFECT: method allows extending a raw-material base, increasing a degree of extraction of immunoglobulins G, A, M and a degree of purification of the end product, reducing end product loss, increasing antibacterial and antiviral activity.

12 cl, 2 ex, 1 dwg

Description

The invention relates to medicine, namely to the production of active substances and medicinal products containing a complex of immunoglobulins G, A, M from human blood plasma.

Human blood plasma contains immunoglobulins of five different classes. Of greatest importance are immunoglobulins of classes G, A, M, which are antibodies and protect the body from various viruses, bacteria and protozoa (for example, chlamydia).

A product containing immunoglobulins can be produced in finished dosage form and used as a medicine (preparation), or as a substance (complex) and used as a component of drugs, including other components.

Various methods are known for isolating a fraction containing immunoglobulins G, A, M. A general approach to the preparation of drugs for the treatment of bacterial and viral infections is the use of so-called precipitation III, or "B", formed in the process of fractionation of human blood plasma with ethanol according to the Cohn method / Aleshkin V.A. et al., 2003 /.

The precipitate of fraction “B” is a pasty mass containing a complex of immunoglobulins, denatured proteins, inactive impurities, lipoproteins and lipids (cholesterol, triglycerides, etc.), a small amount of ethanol. In addition, viruses causing hepatitis, AIDS, and other viral infections may be present in this fraction.

The precipitate of fraction “B” is homogenized in a sodium chloride solution, then treated with an organic solvent (usually chloroform), the proteins are separated from inactive impurities and lipoproteins, and then the target product is isolated by precipitation with polyethylene glycol or concentration.

This scheme for the isolation of the complex of immunoglobulins has several disadvantages associated with the presence in the target product of impurities of polyethylene glycol, fibrinogen, lipoproteins, which complicates sterilizing filtration, reduces the activity of immunoglobulins, increases the loss of the target product and reduces its activity, and also requires immediate drying / RU No. 2199343 / .

Of particular importance is lipids, more precisely cholesterol. Cholesterol (a derivative of fatty acids) is a component of cell membranes and a source of many hormones. Cholesterol is synthesized by the liver, dulls into the general bloodstream and is transferred to various organs and tissues. Due to the fact that cholesterol does not dissolve in aqueous solutions, including in human blood plasma, cholesterol combines with proteins, forming well-soluble complexes - lipoproteins. There are high density lipoproteins and low density lipoproteins. As a result of the action of fractionating agents on the blood plasma and mechanical action, lipoproteins and triglycerides are damaged, lose their solubility and can precipitate, preventing the filtration of protein solutions through membranes.

Various methods have been proposed to address these shortcomings (removal of denatured proteins, lipids, lipoproteins).

According to patent SU 1815820, after purification of fraction “B” with chloroform and dextransulfate, it is recommended that the target product be isolated in the presence of 12% polyethylene glycol.

Patent RU 2060034 for better purification involves the extraction of the alcohol precipitate “B” (Kona fraction) in a forty-sixty-fold volume of sodium chloride solution, followed by removal of the lipoprotein fraction by chloroform.

In order to increase the content of immunoglobulin A in the target product, it was proposed to extract the fraction “B” precipitate first with 10-15% polyethylene glycol, then dissolve the precipitate in distilled water and remove insoluble impurities by centrifugation, and use the supernatant as the target product.

Patent RU 2084229 provides for suspension of precipitate “B” in a 10-fold volume of distilled water, treatment of the solution with chloroform and 0.9% sodium chloride, followed by isolation of the target product with 12% polyethylene glycol.

In order to isolate an IgM-containing concentrate, it is proposed to sequentially treat the precipitate with aerosil and sodium caprylic acid with intermediate centrifugations to remove ballast proteins, and then concentrate / Patent RU 2158136 /.

Patent RU 2189833 provides for sequential treatment of sediment extract “B” with chloroform in a concentration of 0.1 to 3%, copper sulfate in a concentration of 0.1 to 2% at a pH of 6-8, and the target product is isolated by ultrafiltration in the presence of complexing connections. In this case, the content of total lipids in the target product is reduced by 5 times, to 0.308-0.117 g / l.

Patent RU 2199343 provides for the dilution of the “B” precipitate of blood plasma in a saline solution with an ionic strength of 0.01-0.15, precipitation of 13-17% ethanol, suspension of the precipitate and its treatment with 13-15% chloroform, double removal of ballast proteins, additional purification with 4-6% ethanol and isolation of the target product by ethanol precipitation.

Patent RU 2252780 provides for the isolation of the complex of immunoglobulins with ethyl alcohol and 10% chloroform, filtration of a centrifugate with the isolation of the target product with 50% polyethylene glycol, dissolution and sterilizing filtration.

Patent RU 2261112 provides for the extraction of sediment “B” with a 12-fold volume of distilled water, the introduction of sodium chloride to 0.9% concentration, the establishment of pH from 4.0 to 5.0, treatment with 3% chloroform for 14 hours, removing the precipitate, clarifying filtration, isolation of the target product with 12% polyethylene glycol or 26% alcohol, followed by sterilizing filtration, virus inactivation and freeze drying.

Sterilization of the target product can be performed by gamma irradiation with an absorbed dose rate of 3-6 kGy / h, with a total dose of 12-30 kGy before or after packaging.

Despite numerous suggestions for optimizing the scheme for isolating the complex of immunoglobulins, reproduction of the claimed methods in industrial production does not provide the required quality of the target product.

A serious drawback is the complexity of the process, the high cost, the damaging effect of fractionating agents, in particular, 10% chloroform on immunoglobulin molecules, the effect of chloroform vapor on personnel, the decrease in the activity of the target product during storage.

Due to the high content of impurities of fibrinogen and fibrin, coprecipitates with immunoglobulins, the target product is unstable and poorly filtered.

In addition, at most plasma processing plants and blood transfusion stations, only albumin is currently prepared, and the immunoglobulin fraction, the so-called precipitate II + III, or “A”, from which precipitate III, or “B” is obtained from the process of further fractionation. destroy. As a result, the feedstock for the preparation of the complex of immunoglobulins - precipitate III ("B") is a very scarce product.

The closest technical solution is a method for producing an immunoglobulin preparation / Patent RU 2261112, prototype /.

This known method is characterized in that the alcohol precipitate "B" (III Cohn fraction) is extracted with a twelve-fold volume of distilled water, sodium chloride is added to a final concentration of 0.9%, the pH is adjusted from 4.0 to 5.0, chloroform is introduced up to 3% the mixture is kept for 14 hours at 0-3 ° C, centrifuged, the precipitate is removed, clarification filtration is carried out, 50% polyethylene glycol solution up to 12% or ethanol 26% is added to the filtrate, the precipitate is separated by centrifugation, dissolved in 0.3 M glycine with subsequent sterilizing filtration, virus inactivation and freeze-drying.

Testing of this method for producing an immunoglobulin preparation under industrial production conditions showed that under the specified conditions for sediment homogenization (pH from 4.0 to 5.0, sodium chloride 0.9%, chloroform - 3%), the formation of ballast impurities (lipids and lipoproteins) it doesn’t fully occur, and denatured proteins are precipitated with 12% polyethylene glycol or 26% ethanol and end up in the target product, which is very poorly filtered through membranes during clarifying and sterilizing filtration, up to 10-15% of the target product is irretrievably ter etsya. As a result, the product does not possess sufficient antibacterial and antiviral, in particular antichlamydia activity, and storage stability.

The direction of the creation of the drug and the method of its preparation was chosen taking into account the results of studying the causes of the prototype deficiencies - poor extraction of the complex of immunoglobulins from the feedstock, incomplete formation of precipitates of denatured proteins and lipids (cholesterol), which cause loss of the target product and a decrease in its activity during storage.

The technical task of this group of inventions, united by a single technical concept, is to create options for a universal effective way to isolate a complex of immunoglobulins from sediments B (III) or A (II + III), to obtain a drug for the treatment of bacterial and viral infections and to expand the arsenal of these methods and drugs.

The technical result that provides a solution to the problem of this group of inventions consists of:

in expanding the raw material base due to the unlimited use of various types of raw materials - precipitation B (III) or A (II + III);

in increasing the degree of extraction of immunoglobulins G, A, M from raw materials and thereby increasing the yield of the target product;

in reducing losses of the target product and raw materials;

an increase in the degree of purification of the target product due to a decrease in the content of impurities of denatured proteins, lipids and lipoproteins, which is due to an increase in antibacterial and antiviral, in particular antichlamydial, activity by 2-4 times;

in maintaining antibacterial and antiviral activity for 1.5 years when released in the form of a complex of immunoglobulins and maintaining this activity for three years when released in the form of a drug immunoglobulins;

in ensuring the safety of the process by reducing the toxic effects of chloroform on personnel.

The essence of the invention in terms of a method for producing a complex of immunoglobulins of classes G, A, M consists in the fact that as a feedstock, precipitates of B (III), A (II + III) Cohn fractions or a mixture in any ratio that homogenize to 0 are used, 3-0.7% sodium chloride solution to obtain a suspension, which is treated with 1-3% chloroform at a pH of 4.9-5.6, stirred, incubated suspension for 30-60 minutes, centrifuged, and then the target product is isolated by removing excess liquid .

Preferably, a pH of 4.9-5.6 is established by adding a NaOH solution or HCL solution to the suspension, and stirring is carried out at a stirrer speed of 500-1500 rpm, for at least 20 minutes, when the feedstock is homogenized into said sodium chloride solution sodium acetate is introduced at a concentration of 0.01-0.1 M, the selection of the target product is carried out by centrifugation when polyethylene glycol is added using metering devices to a concentration of the last 12% in the mixture, or the isolation of the target product is carried out by ultrafiltration. In addition, before isolating the target product, the stabilizer of the activity of immunoglobulins is introduced into the solution, in particular, polyvinylpyrrolidone at a concentration of 1-5% is introduced into the solution as a stabilizer.

The essence of the invention in terms of a method for producing a preparation containing immunoglobulins of classes G, A, M, consists in the use of precipitates of B (III), A (II + III) Cohn fraction or their mixture in any ratio that homogenize in a 0.3-0.7% solution of sodium chloride to obtain a suspension, which is treated with 1-3% chloroform at a pH of 4.9-5.6, stirred, incubated suspension for 30-60 minutes, centrifuged, and then carry out an additional stage the formation of a precipitate of ballast fractions in a solution with an ionic strength of less than 0.05 when ine pH 4.5 to 5.6 for 12-24 hours followed by removal of an additional centrifugation or filtration and the desired product is then isolated by ultrafiltration.

Preferably, a pH of 4.9-5.6 is established by adding NaOH solution or HCL solution to the suspension, and stirring is carried out at a stirrer speed of 500-1500 rpm, for at least 20 minutes, for better extraction of immunoglobulins during homogenization of the feedstock sodium acetate at a concentration of 0.01-0.1 M is introduced into the indicated sodium chloride solution, the formation of a precipitate of denatured proteins and inactive impurities is carried out in the presence of caprylic acid salts, the target product is poured into bottles and, alternatively, dried from frozen state.

As a result of the method, a complex and / or preparation of immunoglobulins is obtained - an active substance containing immunoglobulins of classes G, A, M, with increased antichlamydia activity, stable for more than 1 year, and after sterilizing filtration, bottling and, as an option, lyophilization, receive a drug for the treatment of bacterial and viral infections with two stages of virus inactivation, which does not contain residual polyethylene glycol, denatured proteins, and the concentration of lipids and lipoproteins (cholesterol) composition less than 0.2 mmol / l.

Marked quality indicators (quantitative content of lipids and lipoproteins, i.e. cholesterol), an immunoglobulin product prepared in accordance with the prototype and variants of the proposed method are shown in table 1.

Table 1 Immunoglobulin preparation Cholesterol, mmol / L Residual Polyethylene Glycol Obtained by the prototype 0.85 + 0.07 0.35% Obtained by the claimed method 0.18 + 0.02 absent

The product obtained by the claimed method (options) contains less cholesterol, and the residual polyethylene glycol is completely absent.

Optimum pH range (4.9 to 5.6) during separation

ballast proteins and lipoproteins with chloroform and an additional step of forming a precipitate of denatured proteins and inactive impurities provide high quality of the target product, free of inactive impurities, as a result of which the target product is easily filtered and losses during clarifying and sterilizing filtration are only 2-3% against 10-15 % of the prototype (see drawing).

The drawing shows the dependence of the filtration rate (vertical) of the target product during clarifying and sterilizing filtration on the pH value (horizontal) at the stage of lipid separation by chloroform.

From the drawing it can be seen that the highest filtration rate of the target product lies in the pH range from 4.9 to 5.6.

The introduction of a stabilizer of polyvinylpyrrolidone into the solution (centrifugate) in an optimal concentration before isolation of the target product ensures that the titer of antibodies to chlamydia is retained for a longer time - for more than 18 months (1.5 years). The minimum titer of antibodies in the complex of immunoglobulins (active substance) containing immunoglobulins of classes G, A, M must be at least 1: 100 and last for more than one year.

Table 2 shows the titer (activity) of antibodies to chlamydia in the complex of immunoglobulins - an active substance containing immunoglobulins of classes G, A, M, prepared according to the prototype and according to the variants of the proposed method

table 2 Upon release In 3 months In 6 months After 9 months After 12 months After 15 months After 18 months Prototype 1: 100 1: 100 1: 100 1: 100 1:50 1:50 1:25 Variants of the proposed method 1: 200 1: 200 1: 200 1: 200 1: 200 1: 100 1: 100

Variants of the method for the production of products containing immunoglobulins are implemented as follows.

Sediment obtained by fractionation of the plasma by the Cohn alcohol method - precipitation B (III), or A (II + III), or a mixture thereof is used as the feedstock. The precipitate is placed in a homogenizer and pour 0.3-0.7% sodium chloride solution in a ratio of 1:10. For better extraction of immunoglobulins, sodium acetate is introduced into the solution at a concentration of 0.01-0.1 M and homogenized for 10-15 minutes. After the disappearance of visible sediment particles using a 1 M NaOH solution or 1 M HCL solution, the pH of the resulting suspension is adjusted to 4.9-5.6 and trichloromethane is added to a concentration of 1-3%. The suspension is mixed at a stirrer speed of 500 - 1500 rpm for at least 20 minutes. The suspension is incubated for 30-60 minutes to inactivate viruses under the influence of chloroform. After that, ballast proteins and lipoproteins are removed by centrifugation. For a more complete removal of lipids, lipoproteins and inactive impurities, an additional purification step is carried out. To do this, reduce the ionic strength in the centrifuge to 0.05 M, set the pH to 4.4-5.6 and leave the solution to form a precipitate for 12-18 hours. For selective denaturation of lipoproteins and better sedimentation, it is advisable to introduce sodium caprylate in a concentration of 0.1-1.0%. Lipoproteins that have lost their solubility in a solution with low ionic strength are removed by centrifugation or filtration. A centrifugate is used to obtain the target product by concentration by ultrafiltration by the interaction of immunoglobulins with polyethylene glycol by adding the latter using dosing devices to a concentration of 12% in the mixture.

In order to increase the activity of the complex of immunoglobulins, a stabilizer, preferably polyvinylpyrrolidone at a concentration of 1-5%, is introduced into the centrifugate before obtaining the desired product.

Get the claimed complex of immunoglobulins containing immunoglobulins of classes G, A, M - an active substance that has increased antichlamydia activity, stable for more than 1 year.

A complex of immunoglobulins containing immunoglobulins of classes G, A, M is subjected to clarifying, sterilizing filtration, standing for 21 days at an acidic pH value (the second stage of inactivation of possibly present viruses), poured into bottles and, as an option, lyophilized. The result is a preparation for the treatment of bacterial and viral infections with two stages of virus inactivation, which does not contain residual polyethylene glycol, denatured proteins, and the concentration of lipids and lipoproteins is less than 0.2 mmol / L.

Example 1

Precipitation of the Cohn fraction A (II + III) and B (III), taken in a ratio of 2: 3, total amount of 15 kg, in portions of 5 kg is placed in a homogenizer, pour 10 times with a volume of 0.3% sodium chloride solution containing 0 , 05 M sodium acetate (50 L per serving) and homogenize the suspension until no visible sediment particles disappear. Then, using a 1 M NaOH solution or 1 M HCL solution, a pH of 4.9 is established and 1.7 L of trichloromethane (chloroform) is added to a final concentration of 3%. Mix the mixture at a speed of rotation of the mixer 500-1500 rpm for 20 minutes. The possible viruses are inactivated, keeping the suspension for 60 minutes and the viruses, lipoproteins and denatured proteins that are possibly present are removed by centrifugation. To obtain a batch of 140 l, the entire procedure with the second and third portions of sediment is repeated. The result is a centrifuge in an amount of 140 liters. An equal volume of a 0.1% sodium caprylate solution is added to the centrifugate, a pH of 4.8 and an ionic strength of less than 0.05 are set and the solution is left to form a precipitate for 18 hours. Lipoproteins that have lost their solubility are removed by centrifugation or filtration. The clarified centrifugate is subjected to concentration by ultrafiltration to a protein content of 5.5-6.5%. A stabilizer, glycine, is introduced to a final concentration of 2%, a pH of 4.7 is established, and it is subjected to clarifying and sterilizing filtration. Get the target product - 19 l of a solution of complex immunoglobulin preparation.

After keeping the product for 21 days (the second stage of inactivation of possibly present viruses), pouring and freeze drying from a frozen state, a preparation is obtained for the treatment of bacterial and viral infections with two stages of virus inactivation, which does not contain residual polyethylene glycol, denatured proteins, with a cholesterol concentration of 0 , 16 mmol / L.

Example 2

Precipitate A (II + III), obtained by fractionation of plasma with the Kohn alcohol method, with a total amount of 13.5 kg, is placed in 4.5 kg portions in a homogenizer, filled with a 10-fold volume of 0.6% sodium chloride solution, (45 l per portion), sodium acetate is introduced at a concentration of 0.01 M and the suspension is homogenized until the visible particles of the precipitate disappear. The pH of the suspension was adjusted to 5.6 and 0.5 L of trichloromethane (chloroform) was added to a concentration of 1%. The suspension is mixed at a stirrer speed of 500-1500 rpm for at least 20 minutes. The suspension is incubated for 30 minutes to inactivate viruses under the influence of chloroform. After that, ballast proteins and lipoproteins are removed by centrifugation.

To obtain a batch of 155 l, the entire procedure for suspension and treatment of the precipitate with chloroform with the second and third portions of the precipitate is repeated. The result is 155 l of a centrifugate, to which a stabilizer, polyvinylpyrrolidone, is added in an amount of 1.6 kg (1%). The centrifuge and the solution of polyethylene glycol are fed to the centrifuge using a metering device until the concentration of the latter is 12% in the mixture.

Get the claimed complex of immunoglobulins containing immunoglobulins of classes G, A, M - an active substance that does not contain residual polyethylene glycol, denatured proteins, and the concentration of lipids and lipoproteins (cholesterol) is less than 0.2 mmol / l, has an increased antichlamydia activity, stable on for 1.5 years.

In accordance with the claimed methods, the experimental production of a medicinal product for the treatment of viral and bacterial infections has been started, an immunoglobulin preparation isolated from human blood plasma with the trade name “Immunoglobulin complex preparation” is used as a medicine, which is a dry mass in a vial containing 300 mg of immunoglobulins G, A, M. The drug is produced according to the pharmacopeia article of the enterprise FSP No. 42-0194-6953-05.

In addition, in accordance with the claimed methods, the experimental production of a medicinal product for the treatment of urogenital chlamydia in women (the drug "Kipferon, suppositories"), which is produced according to FSP 42-0137-0365-00, has been started. As active components (substances), it contains interferon and a complex of immunoglobulins G, A, M, which in this case is a component of the drug. The substance of immunoglobulins is a paste-like thick mass containing a complex of immunoglobulins G, A, M, manufactured according to FSP No. 42-0194-6954-05.

Thus, as a result of the invention, variants of a universal effective method for isolating a complex of immunoglobulins from precipitation B (III) or A (II + III), creating a drug for the treatment of bacterial and viral infections and expanding the arsenal of these methods and preparations are created.

At the same time, the expansion of the raw material base of production, an increase in the degree of extraction of immunoglobulins G, A, M and the degree of purification of the target product, a decrease in the loss of the target product, an increase in antibacterial and antiviral, in particular anti-chlamydial activity by 2-4 times, the preservation of antibacterial and antiviral activity for 1.5 years in the form of a complex of immunoglobulins and maintaining activity for three years in the form of a dry preparation of immunoglobulins, reducing the toxic effect of chloroform on a person .

Literature

1. Sapozhnikova B.C. Development and study of the properties of antistaphylococcal and tetanus toxoid immunoglobulins for intravenous administration.: Author. diss candidate biol. sciences. L., 1990, 20 p.

2. Aleshkin V. A., Lyutov A. G., Afanasyev S. S. and others. Place of immunoglobulin preparations in the treatment and rehabilitation of infectious patients. // New drugs. M., 2003, c. 4, p. 6-32.

3. Rusanov V.M., Skobelev L.I. Fractionation of plasma proteins in the production of blood products. M., Medicine, 1983, p. 75-78.

4. FSP No. 42-0194-6953-05.

5. FSP 42-0504-6737-05.

6. FSP-42-0381-3757-02.

7. SU 1815820 (Aleshkin V.A., Borisova G.V., Kholchev N.V., Ponomareva A.M., Byko-Yanko G.V.)

8. Patent RU 2060034 (Aleshkin V.A., Borisova I.V., Byko-Yanko G.V.)

9. Patent RU 2073525, Aleshkin V.A., Borisova I.V., Novikova L.I.).

10. Patent RU 2084229 (Aleshkin V.A., Borisova I.V., Byko-Yanko G.V.).

11. Patent RU 2158136.

12. Patent RU 2189833 (Zorik A.V., Aleshkin V.A., Lyutov A.G., Borisova I.V.)

13. Patent RU 2199343 (Anastasiev V.V. et al.)

14. Patent RU 2252780 (Avakov A.E., Anastasiev V.V., Piskareva Yu.K.)

15. Patent RU 2261112 (Antonyan R., Bylov K.V.)

16. Patent RU 2255766, Aleshkin V.A., Novikova L.I., Afanasyev S.S., Borisova I.V., Zueva M.M., Zorik A.V.).

17. Patent RU 2261112.

Claims (12)

1. A method of obtaining a product containing immunoglobulins G, A, M, in which precipitation B (III), A (II + III) of the Cohn fraction or a mixture in any ratio that homogenize at 0.3-0 is used as a feedstock , 7% sodium chloride solution to obtain a suspension, which is treated with 1-3% chloroform at a pH of 4.9-5.6, stirred, incubated suspension for 30-60 minutes, centrifuged, and then the desired product is isolated by removing excess liquids. 2. The method according to claim 1, characterized in that the pH value of 4.9-5.6 is established by adding NaOH solution or HCL solution to the suspension, and stirring is carried out at a stirrer speed of 500-1500 rpm for at least 20 minutes . 3. The method according to any one of claims 1 and 2, characterized in that during the homogenization of the feedstock, sodium acetate at a concentration of 0.01-0.1 M is introduced into said sodium chloride solution. 4. The method according to any one of claims 1 and 2, characterized in that the selection of the target product is carried out by centrifugation when polyethylene glycol is added using metering devices to a concentration of the latter 12% in the mixture. 5. The method according to any one of claims 1 and 2, characterized in that the selection of the target product is carried out by ultrafiltration. 6. The method according to any one of claims 1 and 2, characterized in that before the isolation of the target product, a stabilizer of immunoglobulin activity is introduced into the solution. 7. The method according to claim 5, characterized in that, as a stabilizer, polyvinylpyrrolidone is introduced into the solution at a concentration of 1-5%. 8. A method of obtaining a product containing immunoglobulins G, A, M, in which precipitation B (III), A (II + III) of the Cohn fraction or a mixture in any ratio that homogenize at 0.3-0 is used as a feedstock , A 7% solution of sodium chloride to obtain a suspension, which is treated with 1-3% chloroform at a pH of 4.9-5.6, stirred, incubated suspension for 30-60 minutes, centrifuged, and then carry out the stage of additional sedimentation ballast fractions in a solution with an ionic strength of less than 0.05 at a pH from 4.4 to 5.6 for 12-24 h with eduyuschim removal of additional centrifugation or filtration and the desired product is then isolated by ultrafiltration. 9. The method according to claim 8, characterized in that the pH value of 4.9-5.6 is established by adding a NaOH solution or HCL solution to the suspension, and mixing is carried out at a stirrer speed of 500-1500 rpm for at least 20 minutes . 10. The method according to any one of paragraphs.8 and 9, characterized in that for better extraction of immunoglobulins during homogenization of the feedstock, sodium acetate at a concentration of 0.01-0.1 M is introduced into said sodium chloride solution. 11. The method according to any one of paragraphs.8 and 9, characterized in that the formation of a precipitate of denatured proteins and inactive impurities is carried out in the presence of caprylic acid salts. 12. The method according to any one of paragraphs.8 and 9, characterized in that the target product is poured into vials and dried from a frozen state.

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