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Method for removing viruses in preparation of antitoxin and antiserum

An anti-serum and anti-toxin technology, applied in the field of simultaneous removal of residual virus and virus removal, can solve the problems of troublesome removal of organic solvents/detergents, impact on product yield, limited use range, etc., to achieve good stability and ensure High safety and practical value

Inactive Publication Date: 2012-06-27
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

At present, for human blood products, relatively mature virus inactivation / removal methods at home and abroad mainly include physical and chemical methods, of which physical methods include heating, pasteurization, light, nano-membrane filtration, etc., due to most protein It is unstable under the first three conditions, so it should be avoided. Although nano-membrane filtration technology has a good virus removal effect, it is only suitable for proteins with smaller molecules (smaller diameters) due to the limited range of use, and the price is relatively high. expensive and therefore impractical
Chemical methods mainly include: organic solvent / detergent (S / D) method, sodium caprylate method, low pH method, etc. Among them, the S / D method was first used for virus inactivation of human blood products, but it needs 6 The virus can be inactivated within hours, which not only affects the product yield, but also the removal of organic solvents / detergents is troublesome; the sodium octanoate method is a newly developed virus inactivation method, which is safe, fast and efficient; the low pH method is only Suitable for samples that are not sensitive to acids and generally require longer inactivation times

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The method for removing virus in the preparation of tetanus antitoxin comprises the following steps:

[0027] A. After diluting the immune plasma by 2 volumes with water for injection, adjust the pH of the dilution to 2.8 with 2 mol / L HCl;

[0028] B. In every 100ml of the diluent of step A, add 0.1g of gastric enzyme, digest at 29°C for 3 hours to obtain digestive juice;

[0029] C, add the amount of 13g ammonium sulfate per 100ml of digestion solution, in the digestion solution of step B, add ammonium sulfate to form a suspension, and adjust the pH value of the solution with 2mol / L HCl to be 4.8, then add the suspension to After warming to 57°C, keep it for 20 minutes, then lower the temperature to 20°C, and filter the supernatant;

[0030] D. Adjust the pH value of the supernatant in step C to 7.0 with 2 mol / L NaOH, and add 15 g of ammonium sulfate per 100 ml of the supernatant, add ammonium sulfate to the supernatant for precipitation, and let stand for 90 After 1...

Embodiment 2

[0036] The method for removing virus in the preparation of anti-rabies serum comprises the following steps:

[0037] A. After diluting the immune plasma 4 times with water for injection, adjust the pH of the dilution to 3.6 with 2 mol / L HCl;

[0038] B. In every 100ml of the diluent of step A, add 1g of gastric enzyme, digest at 31°C for 1 hour to obtain a digestive juice;

[0039] C, add the amount of 17g ammonium sulfate per 100ml of digestion solution, in the digestion solution of step B, add ammonium sulfate to form a suspension, and adjust the pH value of the solution with 2mol / L NaOH to be 5.6, then add the suspension to After warming to 59 °C, keep it for 30 minutes, then cool down to 35 °C, and filter the supernatant;

[0040]D, with 2mol / L NaOH, adjust the pH value of the supernatant of step C to 7.4, and add the amount of 25g ammonium sulfate per 100ml supernatant, add ammonium sulfate in the supernatant to precipitate, let stand for 180 Minutes later, filter to ge...

Embodiment 3

[0046] The method for removing virus in preparing anti-rabies serum comprises the following steps:

[0047] A. After diluting the immune plasma 3 times with water for injection, adjust the pH value of the diluent to 3.2 with a sodium chloride solution with a mass concentration of 1.0%;

[0048] B. Add 0.5 g of pepsin to every 100 ml of the diluted liquid in step A, and digest at 30°C for 2 hours to obtain a digestive juice;

[0049] C, add the amount of 15g ammonium sulfate according to every 100ml digestion solution, in the digestion solution of step B, add ammonium sulfate to form suspension, and be 1.0% sodium chloride solution with mass concentration to adjust solution pH value to be 5.2, then Heat the suspension to 58°C, keep it for 25 minutes, then cool down to 30°C, filter to get the supernatant;

[0050] D, be 1.0% sodium chloride solution with mass concentration, adjust the supernatant pH value of step C to 7.2, and add the amount of 20g ammonium sulfate by every 100...

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PUM

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Abstract

The invention provides a method for removing viruses in preparation of antitoxin and antiserum. The method comprises the following steps of: diluting immunological blood plasma and adjusting pH value; then digesting by stomach enzyme; heating to denaturize; adsorbing by alum; concentrating / desalinating by ultrafiltration; performing ion exchange chromatography to obtain collected liquid; removing bacteria and filtering; and then subpackaging to obtain an antitoxin and antiserum product. Thus, the viruses which probably exist in raw material serum can be removed simultaneously in the process of preparing the antitoxin and antiserum. The virus removal method for the antitoxin and antiserum provided by the invention has the characteristics of simplicity, quickness economy and practicality; the viruses which probably exist in the raw material serum can be removed completely during production to guarantee the quality of the product.

Description

[0001] technical field [0002] The invention relates to a method for removing viruses, in particular to a method for simultaneously removing residual viruses in the process of preparing antitoxin and antiserum products, and belongs to the field of biotechnology. Background technique [0003] Since 2003, the state has required the biological industry to inactivate / remove viruses from biological extraction products. At present, for human blood products, relatively mature virus inactivation / removal methods at home and abroad mainly include physical and chemical methods, of which physical methods include heating, pasteurization, light, nano-membrane filtration, etc., due to most protein It is unstable under the first three conditions, so it should be avoided. Although nano-membrane filtration technology has a good virus removal effect, it is only suitable for proteins with smaller molecules (smaller diameters) due to the limited range of use, and the price is relatively high. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/16C07K16/06A61K39/395
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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