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Method for inactivating viruses in process of preparing antitoxin and antiserum

An anti-serum and anti-toxin technology, applied in the direction of anti-viral immunoglobulin, serum immunoglobulin, anti-animal/human immunoglobulin, etc. Limit and other problems, to achieve the effect of short processing time, high purity and good safety

Inactive Publication Date: 2012-07-04
玉溪九洲生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the deficiencies such as the long operation time for virus inactivation or removal in existing biological products, the inactivation of active substances is likely to occur, the virus inactivation is not complete, and the scope of application is limited. The present invention provides a simple, fast, economical, and applicable scope It is a method that can simultaneously inactivate the virus existing in the blood during the preparation of antitoxin and antiserum, so as to ensure that antitoxin and antiserum products can be more safely and effectively used in clinical practice

Method used

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  • Method for inactivating viruses in process of preparing antitoxin and antiserum
  • Method for inactivating viruses in process of preparing antitoxin and antiserum
  • Method for inactivating viruses in process of preparing antitoxin and antiserum

Examples

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Embodiment 1

[0030] The method for inactivating virus in preparing tetanus antitoxin comprises the following steps:

[0031] A. After diluting the horse plasma raw material with water for injection to 2 times the volume, adjust the pH value of the diluent to 2.8 with 1N hydrochloric acid solution;

[0032] B. Add 0.1 g of pepsin to every 100 ml of the dilution in step A, and digest at 29°C for 3 hours to obtain a digestive juice;

[0033] C. Add 14g of ammonium sulfate to every 100ml of the digestion solution in step B, and adjust the pH value of the mixture to 4.8 with 1N sodium hydroxide solution, heat the mixture to 57°C and keep it for 30 minutes, then cool down to below 45°C, Separated to obtain supernatant;

[0034] D, the supernatant of step C is diluted to contained protein concentration with water for injection and is 10 g / L, adds medical grade organic solvent tributyl phosphate and medical grade detergent Triton X-100 in this dilution, until Each 100ml of diluent contains 0.25g...

Embodiment 2

[0040] The method for inactivating virus in preparing anti-rabies serum comprises the following steps:

[0041] A. After diluting the plasma raw material with water for injection to 4 times its volume, adjust the pH value of the diluent to 4.0 with 4N hydrochloric acid solution;

[0042]B. Add 1 g of gastric enzyme to every 100 ml of the diluted liquid in step A, and digest at 31°C for 1 hour to obtain the digestive juice;

[0043] C. Add 16g of ammonium sulfate to every 100ml of the digestion solution in step B, and adjust the pH value of the mixture to 5.6 with 4N sodium hydroxide solution, heat the mixture to 59°C and keep it for 30 minutes, then cool down to below 45°C, Separated to obtain supernatant;

[0044] D, the supernatant of step C is diluted to contained protein concentration with water for injection and is 50 g / L, adds medical grade organic solvent tributyl phosphate and medical grade detergent Triton X-100 in this dilution, until Each 100ml of diluent contains...

Embodiment 3

[0050] The method for inactivating virus in preparing antivenom serum comprises the following steps:

[0051] A. After diluting the plasma raw material 3 times with 0.8% NaCl solution, adjust the pH value of the dilution solution to 3.2 with 3N hydrochloric acid solution;

[0052] B. Add 0.5 g of gastric enzyme to every 100 ml of the diluted solution in step A, and digest at 20°C for 22 hours to obtain a digestive solution;

[0053] C. Add 15g of ammonium sulfate to every 100ml of the digestion solution in step B, and adjust the pH value of the mixed solution to 5.0 with 3N sodium hydroxide solution, heat the mixed solution to 57°C and keep it for 30 minutes, then cool down to below 45°C, Separated to obtain supernatant;

[0054] D. Dilute the supernatant of step C with 0.8% NaCl solution to a protein concentration of 25 g / L, and add medical-grade organic solvent tributyl phosphate and medical-grade detergent to the diluted solution Polysorbate-80, until every 100ml of the d...

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Abstract

The invention provides a method for inactivating viruses in the process of preparing antitoxin and antiserum, which comprises the following steps of: after raw material horse plasma is diluted as well as digested, and heated, degenerated and precipitated with gastric enzyme, adding an organic solvent and a decontaminating agent to carry out virus inactivation; carrying out ammonium sulfate separation, alum adsorption, ultrafiltration concentration / desalination and ion-exchange column chromatography to obtain concentration solution; and after sterilization and filtration, subpacking to obtain a product. The method has the beneficial effects that lipid envelopes of the viruses are damaged by the organic solvent and the decontaminating agent, so that the infectivity of the viruses is lost, thereby the viruses remaining in the plasma are completely inactivated; meanwhile, the structure and the function of protein are kept, and then the residues of the organic solvent and the decontaminating agent are effectively removed, so that the purity of the product is further improved; the method has the advantages of simplicity in virus inactivation operation, short treatment time and good economy and practicability; and moreover, the prepared product is high in purity and good in safety.

Description

technical field [0001] The invention relates to a method for simultaneously inactivating viruses in the process of preparing antitoxin and antiserum, and belongs to the field of biotechnology. Background technique [0002] Antitoxin and antiserum (Antitoxin / Antiserum) refer to the use of specific antigens to immunize animals, then collect high-titer immune serum, and after extraction and purification, prepare antibodies containing intact antibodies (IgG) or antibody fragments [F(ab') 2 ] immunoglobulin products. For the prevention and treatment of infections caused by the corresponding diseases. [0003] Because the raw materials of antitoxin and antiserum come from horses, and horses are raised in an open environment, they may carry more viruses, so the products have biological safety problems. In order to ensure the safety of this product, in addition to controlling the breeding environment of immunized horses and the quality of raw plasma, how to effectively remove or i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/06C07K16/12C07K16/10C07K16/18C07K1/36C07K1/34C07K1/30C07K1/18
Inventor 吴笛杨冬罗靖雄
Owner 玉溪九洲生物技术有限责任公司
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