RU2470664C2 - Method for producing immunoglobulin for intravenous introduction of immunoglobulin m enriched preparation, and preparation prepared by such method - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine. A preparation is produced of donor blood plasma by protein fractionation in ethanol at temperature below 0°C (Cohn method); the produced filtrate B (III) is used to recover purified immunoglobulin G; the produced deposition B (III) is used to recover a purified immunoglobulin M enriched concentrate; it is followed by separate decrease of anti-complement activity of immunoglobulin G and immunoglobulin M; then a pharmaceutical composition is prepared by combining immunoglobulins G and M with decreased anti-complement activity by mixing immunoglobulins G and M taken in equal proportions. Decrease of anti-complement activity of immunoglobulins G and M is differentiated. For immunoglobulin G - to the value max. 1 hemolytic unit (CH50) per 1 mg of immunoglobulin by molecular filtration dialysis with the use of purified water at pH 3.8-4.5; for immunoglobulin M to the value max. 1 hemolytic unit (CH50) per 1 mg of immunoglobulin with keeping a semi-finished product at temperature 45±5°C for 24-48 hours. The preparation is to be virus-inactivated and dried.

EFFECT: group of invention provides producing the immunoglobulin M enriched preparation with higher IgG biological activity and lower spontaneous anti-complement activity.

8 cl, 2 ex, 1 tbl

Description

The invention relates to medicine, namely to the production of an immunoglobulin preparation enriched with immunoglobulin M (IgM) and suitable for intravenous administration.

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

For the treatment of severe bacterial and viral infections, intravenous administration of immunoglobulins, which are isolated from blood plasma and purified immunoglobulin G (IgG), is currently widely and successfully used. The choice of therapeutic agents containing IgG is quite large: they all differ in their quality, frequency of adverse reactions, form, physical parameters, duration of circulation in the body and effectiveness. / Sapozhnikova B.C., 1990, Berkman S.A. et al., 1990 /.

Commercial immunoglobulin preparations are 95% IgG. In especially severe septic conditions, it is proposed to use the intravenous administration of a drug enriched with IgM.

It is believed that IgM, which is part of the composition of the drug, is highly effective in binding bacterial cells and their toxins.

The difficulty in developing such a drug lies in the difficulty in ensuring a high yield of IgM from the feedstock, as well as in that IgM has a particularly high complement binding activity in vitro and special conditions are necessary to eliminate the spontaneous ability of the drug to activate complement.

Immunoglobulin M (IgM) can be isolated from plasma by treatment with hexane, precipitation with 6% polyethylene glycol 6000 (PEG) and chromatography on 6% imidus agarose / patent RU 1732518 /.

A known method of obtaining a drug for intravenous administration with an IgM content of more than 5 wt.% By treating the plasma fraction containing IgM with a protease / patent RU 2191032 /. Obtaining an IgM-containing concentrate is possible by sequentially treating sediment B (III) with aerosil and sodium caprylic acid with intermediate centrifugations to remove ballast proteins, followed by concentration / patent RU 2158136 /.

An immunoglobulin preparation containing IgM and IgA is obtained by dissolving precipitate B in 0.1 M acetate buffer, treated with octanoic acid, dialyzed against a piperazine and sodium chloride solution, and then conduct ion exchange chromatography / patent US 4256631 /. You can use a non-toxic polymer - polyethylene glycol / patent US 3808189 /.

The so-called IgGAM preparation can be isolated from plasma or precipitate B using a combination of precipitation methods with caprylic acid and ethanol / Stembuch M., 1980 /.

Obtaining an IgM enriched immunoglobulin fraction is not enough. One of the main indicators of the quality of intravenous immunoglobulin preparations is the so-called anti-complementary activity - the ability, after intravenous administration, to spontaneously bind and activate complement. Activation of complement (whey protein system) leads to the release of vasoactive substances that cause a drop in blood pressure, the appearance of shortness of breath, tremors, nausea, and in some cases collapse / Barandun S. et al., 1962 /.

In this regard, one of the most important stages of the technological process of obtaining an immunoglobulin preparation is the elimination or reduction of anticomplementary activity.

Several methods are known for reducing the anti-complementary activity of immunoglobulins: treatment with proteolytic enzymes / SU patent No. 618958 /, modification of immunoglobulin by acetylation / US patent No. 4356173 / or sulfonation / US patent No. 4360457 /, aging at 37 ° C for 18 hours / Mostovskaya E .V., 2004 /, incubation at a temperature of 30-45 ° C / EP No. 0085747 /, gel treatment with aluminum hydroxide followed by thermal treatment / Zubkova N.V., 2002 /.

The described methods effectively eliminate the anti-complementary activity of preparations containing IgG. The spontaneous anticomplementary activity of IgM is much higher due to the fact that the IgM molecule consists of five monomers that actively bind complement. Therefore, to reduce the anti-complementary activity of an IgM-enriched preparation, tough methods are used - treatment with betapropiolactone, sulfonation, or heating.

Thus, US patent 4318902 provides for the treatment of a 5% solution of immunoglobulin containing IgM with betapropiolactone in an amount of 0.05-0.15 ml per 100 ml of immunoglobulin at a temperature of 20-37 ° C for 4-6 hours. This method is the basis of the production technology of the drug "Pentaglobin", currently produced by the company "Biotest" / Stephan W., 1983 /. In this case, the preservation of the biological activity of the drug is postulated, which can be considered reliable only with respect to the IgM component, but unrealistic with respect to the IgG component due to the high rigidity of this treatment.

EP 0450412 proposes to process an IgM-containing concentrate having a pH of from 4.0 to 5.0 at a temperature of from 40 to 62 ° C. for more than 10 minutes.

A preparation containing more than 5% IgM and more than 10% IgA can be obtained by treating fraction A (II + III) or B (III) with octanoic acid, purification by anion exchange chromatography, and processing the eluate with a pH value of 4.0-4.5 at a temperature of 50 -54 ° C for from 1 minute to 24 hours / US patent 5075425, US No. 5410025 /.

The disadvantage of all the described methods for producing preparations of immunoglobulin enriched with IgM is the fact that in order to eliminate the anti-complementary activity of IgM, the complex preparation of IgGAM enriched with IgM is rigidly processed, which leads to a significant decrease in the biological activity of IgG, since its molecule consists from only one monomer, which has 5 times less ability to bind complement and is much more sensitive to the damaging effects of chemical or temperature effects, it IgM molecule.

The closest technical solution is a method for producing an immunoglobulin preparation for intravenous administration and a preparation for intravenous administration. This known method for the preparation of an immunoglobulin preparation for intravenous administration involves obtaining a fraction containing a complex of immunoglobulins G, A, M from sediment B (III) and treating a 5% immunoglobulin solution containing IgM with betapropiolactone at a temperature of 20-37 ° C for 4-6 hours. This solution is used in the production technology of the commercial IVIG drug enriched with IgM-Pentaglobin (US 4318902, prototype).

Its disadvantages are: low activity of antibodies to various antigens (Mostovskaya EV, 2004); high residual anticomplementary activity, requiring very slow intravenous administration of the drug; low ability to activate macrophages, since a significant part of IgG Fc fragments is damaged during harsh chemical treatment with beta-propiolactone.

The direction of creation of the claimed drug and the proposed method for its preparation was chosen taking into account the different anti-complementary activity of IgM and IgG and their unequal resistance to chemical and physical factors.

The technical task of this group of inventions, united by a single inventive concept, is to create a method for producing an effective immunoglobulin preparation for intravenous administration enriched with IgM, as well as to create an effective immunoglobulin preparation for intravenous administration enriched with IgM, while fully preserving the biological activity of IgG.

The technical result that provides the solution of the problem is to reduce the anti-complementary activity of IgG and IgM included in the pharmaceutical composition while maintaining their biological activity, i.e. preservation of all biological properties of IgG while at the same time effectively reducing the anti-complementary activity of IgM, increasing the therapeutic activity of the pharmaceutical composition (preparation).

The essence of the invention in terms of the method is that to obtain an immunoglobulin preparation for intravenous administration, donor blood plasma is subjected to protein fractionation with ethanol at a temperature below 0 ° C (Kohn's method), purified immunoglobulin G is isolated from the obtained filtrate B (III), from the obtained precipitate B (III), a purified concentrate enriched with immunoglobulin M is isolated, after which individually the anti-complementary activity of immunoglobulin G and the anti-complementary activity of the immunoglobe are individually reduced lin M, and then make a pharmaceutical composition by a compound of immunoglobulins G and M with low anticomplementary activity.

Preferably, the preparation is made by mixing in equal proportions of immunoglobulins G and M with reduced anticomplementary activity.

In this case, the decrease in the anticomplementary activity of immunoglobulins G and M is carried out differentially by different methods, selected on the basis of more complete conservation of the biological activity of each of the immunoglobulins G and M under the influence of physical and chemical factors of the corresponding method.

The anti-complementary activity of immunoglobulin G is reduced to a value of not more than 1 hemolytic unit (CH50) per 1 mg of immunoglobulin by molecular dialysis filtration using purified water with a pH of 3.8-4.5, and the anti-complementary activity of immunoglobulin M is reduced to a value of not more than 1 hemolytic unit (CH50) per 1 mg of immunoglobulin, maintaining the semi-finished product at a temperature of 45 ± 5 ° C for 24-48 hours.

The essence of the invention in terms of the preparation lies in the fact that the preparation of an immunoglobulin for intravenous administration contains immunoglobulins G and M with reduced anticomplementary activity obtained by the above method, in order to obtain an immunoglobulin preparation for intravenous administration, donor blood plasma is subjected to protein fractionation with ethanol at a temperature below 0 ° C Cohn method), purified immunoglobulin G is isolated from the obtained filtrate B (III), purified concentrate enriched from the obtained precipitate B (III) is isolated immunoglobulin M, after which individually the anti-complementary activity of immunoglobulin G is reduced and the anti-complementary activity of immunoglobulin M is reduced, and then the pharmaceutical composition is made by combining immunoglobulins G and M with reduced anti-complementary activity. Preferably, the preparation is made by mixing in equal proportions of immunoglobulins G and M with reduced anticomplementary activity. In this case, the decrease in the anticomplementary activity of immunoglobulins G and M is carried out differentially by different methods, selected on the basis of more complete conservation of the biological activity of each of the immunoglobulins G and M under the influence of physical and chemical factors of the corresponding method. The anti-complementary activity of immunoglobulin G is reduced to a value of not more than 1 hemolytic unit (CH50) per 1 mg of immunoglobulin by molecular dialysis filtration using purified water with a pH of 3.8-4.5, and the anti-complementary activity of immunoglobulin M is reduced to a value of not more than 1 hemolytic unit (CH50) per 1 mg of immunoglobulin, maintaining the semi-finished product at a temperature of 45 ± 5 ° C for 24-48 hours.

Preferably, the preparation contains in equal proportions immunoglobulins G and M with reduced anti-complementary activity and a stabilizer, preferably maltose, in a concentration of not more than 10%.

The drug is subjected to virus inactivation and drying.

The method is as follows.

Each batch of raw materials for the preparation of the drug is prepared from blood plasma obtained from a group of donors by fractionating its proteins with ethanol at a temperature below 0 ° C (Kohn's method).

Purified immunoglobulin G is isolated from precipitate A (II + III) by one of the known methods. IgM enriched concentrate is isolated from precipitate B (III). Two semi-finished products are obtained, one of which is an immunoglobulin G solution, and the second semi-finished product is an IgM enriched immunoglobulin solution. Note: each of these semi-finished products also contains IgA, to simplify the description reflects the sequence of actions on the obtained semi-finished products in relation to their main active components: IgM and IgG.

The anti-complementary activity of immunoglobulin G is reduced by one of the known methods (preferably, in accordance with patent No. 2302427). Then, the anti-complementary activity of immunoglobulin M is reduced by one of the known methods. For example, a semi-finished product enriched with immunoglobulin M is maintained at a temperature of 45 ± 5 ° C for 24-48 hours.

Thus, the method for producing immunoglobulin enriched with IgM suitable for intravenous administration is characterized in that the decrease in the anti-complementary activity of IgG and IgM included in the preparation is carried out differentially (separately reduce the anti-complementary activity of the semi-finished product enriched with IgM, separately the semi-finished product containing IgG) with taking into account their resistance to physical and chemical factors on the basis of ensuring the most complete conservation of the biological activity of immunoglobulins, after which they make up the farm tsevticheskuyu composition.

In conclusion, they formulate a pharmaceutical composition comprising preferably 1 part of a semi-finished product of immunoglobulin G with reduced anti-complementary activity and preferably 1 part of a semi-finished product enriched in immunoglobulin M with reduced anti-complementary activity. The result is a preparation enriched in immunoglobulin M and suitable for intravenous administration.

Example 1

The raw material for obtaining the drug is quarantined blood plasma from a group of donors by fractionation of its proteins with ethanol at a temperature below 0 ° C (Kohn's method).

In fraction III (filtrate "B") of the Cohn alcohol method, a pH of 3.5–5.0 was established and concentrated on an ultrafiltration unit to reduce the solution volume by 10–12 times. Then ethanol is removed using 5-6 volumes of purified water with a pH value of 3.8 to 4.5, and then the immunoglobulin solution is concentrated to a protein content of 6-7%. After this, the hydration shells of the immunoglobulin are restored, for which a low molecular weight substance, preferably maltose or NaCL, is introduced into the immunoglobulin solution in an amount of 0.1-2.5% and molecular diafiltration is carried out using purified water with a pH value of 3.8-4.5 . A semi-finished product of immunoglobulin G with low anti-complementary activity is obtained.

A complex of immunoglobulins (G + A + M) is extracted from the precipitation of fraction B (III). The precipitates are homogenized in a 0.3-0.7% sodium chloride solution to obtain a suspension that is treated with 1-3% chloroform at a pH of 4.9-5.6, stirred, the suspension is kept for 30-60 minutes, centrifuged, and then isolated target product by removing excess fluid. A stabilizer is introduced, preferably maltose, at a concentration of 10%, and the anti-complementary activity of immunoglobulin M is reduced, keeping the semi-finished product at a temperature of 45 ± 5 ° C for 24-48 hours.

In conclusion, a pharmaceutical composition is made up of 1 part of a semi-finished product of immunoglobulin G with low anti-complementary activity and 1 part of a semi-finished product enriched in immunoglobulin M with reduced anti-complementary activity. The result is a pharmaceutical composition - the target product, which is a complex of immunoglobulins (G + A + M), suitable for intravenous administration with high biological activity.

Moreover, an immunoglobulin preparation for intravenous administration enriched with IgM contains not less than 50% IgG, not less than 5% IgM, preferably from 5% to 25% IgM, stabilizer and sodium chloride not more than 0.4% with anticomplementary activity less than 1 unit of CH 50 per 1 mg of protein.

The preparation contains a stabilizer and sodium chloride not more than 0.4%. The drug is suitable for intravenous administration and includes immunoglobulin G of at least 50%, immunoglobulin M of at least 5% (the rest is IgA and other proteins), and consists of immunoglobulin G with reduced hydration membranes and immunoglobulin M with reduced anticomplementary activity.

Example 2

The initial raw material for the preparation of the drug is blood plasma of donors, which was quarantined and checked for the absence of markers of pathogens of vector-borne diseases. Plasma is fractionated with ethanol at a temperature below 0 ° C (Kohn's method).

The supernatant of fraction B (III) in an amount of 300 L was placed in a reactor (vessel) and a pH value of 4.1 was established using a 1 M HCL solution, after which it was concentrated in an ultrafiltration unit. After reducing the solution to 30 L, ethanol is removed using 150 L of purified water with a pH of 4.2, and the immunoglobulin is concentrated to a volume of 25 L. The hydration shells of the immunoglobulin are restored by molecular filtration. After that, the pH is checked and, if necessary, adjusted, the stabilizer is introduced and the protein content (concentration) is set to 5.2%. After clarifying and sterilizing filtration, a semi-finished product of immunoglobulin G with anticomplementary activity of 10 mg of protein that does not activate 2 CH _{50 is} obtained.

To obtain a semi-finished product containing IgM, a precipitate of fraction B (III) in an amount of 15 kg is placed in a homogenizer, filled with a 10-fold volume of a 0.3% sodium chloride solution containing 0.05 M sodium acetate, and the suspension is homogenized until the visible particles of the precipitate disappear . Then, using a 1 M NaOH solution or 1 M HCL solution, a pH of 4.9 is established and trichloromethane is added to a final concentration of 1.5%. Mix the mixture at a speed of rotation of the mixer 500-1500 rpm for 20 minutes. Inactivation of possibly present viruses is carried out, keeping the suspension for 60 minutes, and possibly present viruses, lipoproteins and denatured proteins are removed by centrifugation. An equal volume of a 0.1% sodium caprylate solution is added to the centrifugate, a pH of 4.8 is set, the ionic strength is less than 0.05, and the solution is left to form a precipitate for 18 hours. The precipitate is removed, and the clarified centrifugate is subjected to concentration by ultrafiltration to a protein content of 5.5-6.5%. A stabilizer, maltose, is introduced, a pH of 4.2 is established, and it is clarified and sterilized by filtration. Get the semi-finished product containing 18% immunoglobulin M in an amount of 19 l

The anti-complementary activity of immunoglobulin M is reduced, keeping the semi-finished product at a temperature of 50 ° C for 36 hours.

After that, a pharmaceutical composition is made up by combining one part of a semi-finished product of immunoglobulin G and 1.5 parts of a semi-finished product enriched with immunoglobulin M. The result is a target product containing a complex of immunoglobulins G (78.4%), M (11.8%) and A ( 9.8%). The pharmaceutical composition is re-clarified and sterilized by filtration, virus inactivation, controlled, poured into vials to obtain an immunoglobulin preparation enriched with immunoglobulin M suitable for intravenous administration containing immunoglobulin with increased biological activity of IgG and low spontaneous anticomplementary activity.

The high biological activity of the drug obtained according to this group of inventions, with reduced to less than 1 unit of CH50 per 1 mg of protein of the spontaneous ability of the drug to activate complement, is confirmed by comparison with the prototype (pentaglobin).

A comparative study of the effect of various preparations of intravenous immunoglobulins (IVIG) of domestic and foreign production on the oxidase activity of neutrophils in the test of luminol-dependent chemiluminescence (LHCL) in vitro in healthy donors was carried out. For this, a leukocyte suspension of 10 healthy donors was individually incubated with each of the IVIG containing IgG: Gabryglobin-IgG (Ivanovo), Normal human immunoglobulin (ICH) (N. Novgorod), Humaglobin (Hungary), Octagam (Austria), ICH Vienna (Italy) ), Intraglobin (Germany) and IVIG enriched with IgM: Pentaglobin (Germany) and the experimental preparation Gabriglobin-IgM (the claimed preparation), evaluating the oxidase activity by the LHLC method using a LUCY-2 luminometer (probe-luminol, respiratory explosion activator) "- zymosan (" Sigma ", CI UA). We took into account the stimulation index (IS) - the ratio of LHCH with IVIG to LHCL without IVIG; IS> 1.1 was taken as normal.The results of the study showed that LHCL in 10 donors averaged 13.2 ± 3.1 mV / min, and LZHL induced by zymosan - 146.8 ± 13.4 mV / min (p <0.05). ICH, Octagam, Intraglobin showed oxidase activity in 50% of donors (IP 1.25 ± 0.10); Pentaglobin - in 60% (IP 2.04 ± 0.42); Gabryglobin-IgM - in 70% (IP 1.33 ± 0.16); Gabriglobin-IgG, Humaglobin, ICH Vienna - in 80% (IP 1.54 ± 0.40). The data obtained demonstrated the uneven effect of different IVIG on the oxidase activity of neutrophils in donors (p <0.05), which is important for the clinical use of IVIG. Among domestic IVIG, the greatest effect was found in the drug Gabrilobin-IgG and Gabrilobin-IgM / Kozlova M.N. et al., 2008 /.

Among the many aspects of the action of intravenous immunoglobulin, the main factor determining the effectiveness of the drug is the neutralization of microbial toxins, viruses and the binding of pathogenic antibodies. This is because, according to Good R.A., Lorenz E. (1991), 1 g of normal intravenous immunoglobulin contains more than 10,000,000 antibody molecules of various specificity.

In the process of obtaining a pharmaceutical preparation, antibodies - immunoglobulins in the process of fractionation and concentration lose their native ability and are damaged due to exposure to ethanol, chemical agents and the removal of anticomplementary activity.

Therefore, the titer of antibodies to various viruses and bacteria is one of the most important quality parameters of intravenous immunoglobulins.

An enzyme immunoassay was used to study the content of antibodies to the causative agents of certain infectious diseases in the prototype (Pentaglobin) and the preparation obtained by the claimed method (Gabriglobin-IgM).

Used enzyme-linked immunosorbent test systems manufactured by Vector-Best, the results were expressed as titer (last positive dilution of immunoglobulin) or arbitration units AE / Handsfield H. et al., 1987 /. The level of IgG antibodies to the polysaccharide of meningococcus was presented as a "Positivity Ratio", calculated as the OD: cut off. Antibodies to pertussis, diphtheria toxins, and meningococcus were detected by diagnostic enzyme-linked immunosorbent assay systems developed by the employees of the Federal State Institution of Medical Research, Moscow. G.N.Gabrichevsky "/ Makarova S.I., 2005 /.

The data are presented in the table.

The content of antibodies to causative agents of certain infectious diseases in immunoglobulin preparations No. p / p Name of antibodies The content of antibodies in IG preparations obtained: prototype M ± m the claimed method, M ± m one. AT measles virus, AE 2869 ± 332 8834 ± 721 2. Anti-diphtheria antibodies, mcg / ml 27.9 ± 3.6 36.2 ± 5.4 3. Antitussive antibodies, U / ml 72 ± 7.9 279.4 ± 23.1 four. AT polysaccharide meningococcus serotype A, KP 2.79 ± 0.16 2.91 ± 0.11 5. AT to the herpes simplex virus generic antigen 16 240 ± 1195 24862 ± 1280 6. AT to cytomegalovirus, AE 2188 ± 92 3575 ± 172 7. AT to Varicella Zoster virus, AE 443 ± 40 360 ± 20 8. Herpes simplex virus AT, serotype 2 348 ± 83 5636 ± 1264 9. Herpes simplex virus AT, serotype 6 921 ± 2 1166 ± 19 10. Rubella virus ATE 5524 ± 274 11350 ± 2762

The results indicate that the titer of antibodies to the vast majority of antigens was higher in the preparation obtained by the claimed method.

Thus, as a result of the implementation of the present group of inventions, an immunoglobulin preparation enriched with immunoglobulin M suitable for intravenous administration is obtained containing immunoglobulin with increased biological activity of IgG and low spontaneous anticomplementary activity of the pharmaceutical composition.

Information sources

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

2. Berkman S.A., Lee M.L., Gale R.P. Clinical uses of intravenous immunoglobulins.// Annals of internal medicine. - 1990. - V.112, No. 4. - P.278-292.

3. Patent RU 1732518.

4. Patent RU 2191032.

5. Patent RU 2158136.

6. Patent US 4256631.

7. Patent US 3808189.

8. Steinbuch M. Protein fractionation by ammonium sulphate, rivanol and caprylic acid precipitation. In Methods of plasma protein fractionation. Ed. Curling J.M. Academic Press, N.Y., 1980.

9. Barandun S., Kistler P., Jenet F., Isliker H. Intravenous administration of human γ-globulin. // Vox sanguinis. - 1962. - V.7. - No. 2. - P.157-174.

10. Patent SU No. 618958.

11. US patent No. 4356173.

12. US patent No. 4360457. Ono et al. S-sulfonated immunoglobulin composition having a high monomer content and a process for production therefore. 1982.

13. Mostovskaya E.V. Optimization of the technology for producing a liquid form of immunoglobulin for intravenous administration. Abstract. diss. Cand. M., 2004, 22 p.

14. EP No. 0085747.

15.3ubkova N.V. Physico-chemical and biological properties of immunoglobulin preparations with different methods for the removal and inactivation of viruses. Abstract. diss. Cand. biol. sciences. M., 2002, 24 p.

16. US 4,318,902.

17. Stephan W. Intravenous IgM by means of treatment of Cohn fraction III with β-propiolactone. In: Curling J.M. (ed) Pharmacia fine chemicals. Uppsala, Budapest, 1982, P.153.

18. EP 0450412.

19. Stephan W., Spilles C., 1983.

20. US patent 5075425. Kotitschke R., Stephan W., Moller W. et al. Process for the preparation of pharmaceutical which contains IgG, IgA and IgM and can be administered intravenous. 1990.

21. US patent 5410025. Moller W., Piechaczek. Unmodified intravenously administered immunoglobulin preparations containing imminoglobulin M and / or A., 1995.

22. Kozlova M.N. et al. Russian immunological journal, 2008, Vol. 2, No. 2-3, p.142.

23. Good R.A., Lorenz E. Historic aspects of intravenous immunoglobulin therapy. Cancer., 1991, N 68, 6 Suppl., P.1415-1421.

24. Handsfield H.H., Wandell M., Golgstein L., Shriver K. // Screening and diagnostic performance of enzyme immunoassay for antibody to lymphadenopathy-associated virus. Journal of clinical microbiology., 1987. - N.5, p. 879-884.

25. Makarova S.I. Identification of anti-diphtheria antibacterial antibodies using enzyme-linked immunosorbent assay. Abstract. diss. Cand. biol. Sciences, M., 2005, 22 p.

Claims (8)

1. A method for preparing an immunoglobulin preparation for intravenous administration, in which donor blood plasma is subjected to ethanol fractionation of proteins at a temperature below 0 ° C (Kohn's method), purified immunoglobulin G is isolated from the obtained filtrate B (III), and precipitate B (III) is isolated purified concentrate enriched with immunoglobulin M, after which individually lowering the anti-complementary activity of immunoglobulin G and reducing the anti-complementary activity of immunoglobulin M are carried out separately, and then they are pharmaceutical w composition by coupling of immunoglobulins G and M with low anticomplementary activity. 2. The method according to claim 1, characterized in that the preparation is made by mixing in equal proportions of immunoglobulins G and M with reduced anti-complementary activity. 3. The method according to any one of claims 1 and 2, characterized in that the decrease in the anti-complementary activity of the immunoglobulins G and M is carried out differentially, using different methods selected based on more complete conservation of the biological activity of each of the immunoglobulins G and M under the influence of physical and chemical factors corresponding method. 4. The method according to claim 3, characterized in that the anti-complementary activity of immunoglobulin G is reduced to a value of not more than 1 hemolytic unit (CH50) per 1 mg of immunoglobulin by molecular dialysis filtration using purified water with a pH value of 3.8-4, 5. 5. The method according to claim 3, characterized in that the anti-complementary activity of the immunoglobulin M is reduced to a value of not more than 1 hemolytic unit (CH50) per 1 mg of immunoglobulin, maintaining the semi-finished product at a temperature of 45 ± 5 ° C for 24-48 hours 6. An immunoglobulin preparation for intravenous administration containing immunoglobulins G and M with reduced anti-complementary activity, obtained by the method according to claim 1. 7. The drug according to claim 6, characterized in that it contains in equal proportions immunoglobulins G and M with reduced anti-complementary activity and a stabilizer, preferably maltose, in a concentration of not more than 10%. 8. The drug according to any one of paragraphs.6 and 7, characterized in that it is subjected to virus inactivation and drying.