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Method for separating and purifying anti-hepatitis B immune globulin from plasma or plasma component

An immunoglobulin, separation and purification technology, applied in the direction of serum immunoglobulin, antibody, digestive system, etc., can solve the problems of waste of plasma raw materials and reduced utilization of plasma resources, and achieve high affinity, high purity and antibody titer. , good safety effect

Inactive Publication Date: 2011-07-20
许贤豪
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This makes a large number of plasma raw materials whose potency is lower than 8IU / ml but significantly higher than that of ordinary plasma cannot be used for the preparation of anti-hepatitis B immunoglobulin, resulting in the waste of plasma raw materials and greatly reducing the utilization rate of plasma resources.

Method used

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  • Method for separating and purifying anti-hepatitis B immune globulin from plasma or plasma component
  • Method for separating and purifying anti-hepatitis B immune globulin from plasma or plasma component

Examples

Experimental program
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Embodiment 1

[0022] The preparation of embodiment 1 affinity chromatography medium

[0023] The matrix of the chromatographic medium can be dextran, agarose, or other porous polysaccharide polymer hydrophilic substances; or a medium with porous rigid support and the surface covered with polysaccharide polymer hydrophilic substances. By controlling the chemical reaction conditions, the hydroxyl groups on the substrate are chemically modified to introduce highly reactive small molecule chemical groups such as cyanogen bromide, epoxy groups or mercapto groups. After washing to remove unreacted small molecule chemical groups and stabilizing reactive groups, the chemical activation of the chromatographic medium matrix is ​​completed.

[0024] The hepatitis B surface antigen (HbsAg) material containing antigenic determinants is derived from the genetic engineering recombination method, that is, obtained from the hepatitis B vaccine manufacturer or the hepatitis B surface antigen reagent manufact...

Embodiment 2

[0025] Example 2 Component II affinity chromatography separation of anti-hepatitis B immunoglobulin

[0026] 100 ml of the component II sample with a protein content of 18.2% was added to 100 ml of sodium dihydrogen phosphate / disodium hydrogen phosphate buffer solution with pH 7.4 to dilute evenly, and used as the sample before affinity chromatography purification. 50ml of genetically engineered HBsAg affinity chromatography column was equilibrated with sodium dihydrogen phosphate / disodium hydrogen phosphate buffer solution of pH 7.4 for 5 column volumes, loaded with 200ml of component II dilution, and collected 300ml of the breakthrough peak. After loading the sample, rinse with sodium dihydrogen phosphate / disodium hydrogen phosphate buffer solution to the baseline. The anti-hepatitis B immunoglobulin was eluted with a glycine / hydrochloric acid buffer solution of pH 2.5, and 80 ml of the eluted anti-hepatitis B immunoglobulin component peak was collected.

[0027]As can be s...

Embodiment 3

[0032] Example 3 Separation of anti-hepatitis B immunoglobulin by plasma affinity chromatography

[0033] 2000ml of plasma sample with anti-hepatitis B activity of 8IU / ml was added to 2000ml pH7.4 sodium dihydrogen phosphate / disodium hydrogen phosphate buffer solution to dilute evenly, and used as the sample before affinity chromatography purification. 50ml of genetically engineered HBsAg affinity chromatography column was equilibrated with pH 7.4 sodium dihydrogen phosphate / disodium hydrogen phosphate buffer solution for 5 column volumes, loaded plasma sample dilution 4000ml, and collected breakthrough peak 4150ml. After loading the sample, rinse with sodium dihydrogen phosphate / disodium hydrogen phosphate buffer solution to the baseline. The anti-hepatitis B immunoglobulin was eluted with a pH 2.5 glycine / hydrochloric acid buffer solution, and 94 ml of the eluted anti-hepatitis B immunoglobulin component peak was collected.

[0034] From the detection results of hepatitis B...

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Abstract

The invention discloses a method to separate and purify Anti-hepatitis B virus Immunoglobulin from blood plasmas or blood plasma components. The method is to sieve blood plasmas of anti-hepatitis B virus Immunoglobulin with the titer no less than 2IU / ml and mix the blood plasmas. Small molecular chemical groups with high reaction activity are introduced to the hydroxyl group of the hydroxyl of high molecular hydrophibic substances of polysaccharose, and chromatography base soliquoid is acquired. The Hepatitis B surface antigen solution with antigen determinants is added into the chromatography base soliquoid to be coupled and acquire affinity chromatography media; the components containing Immunoglobulin C that acquired from mixed blood plasmas through low temperature ethanol technique, or blood plasmas are directly used as material of purified Anti-hepatitis B virus Immunoglobulin; affinity chromatography media is used for collecting the components of Anti-hepatitis B virus Immunoglobulin; the purified Anti-hepatitis B virus Immunoglobulin is concentrated and inactivated by steps, added with stabilizer and packed, finally the medical preparation of Anti-hepatitis B virus Immunoglobulin is acquired. The recycling ratio of Anti-hepatitis B virus Immunoglobulin of the method is near 100 percent.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for separating and purifying anti-hepatitis B immunoglobulin from plasma or plasma components. Background technique [0002] Chronic hepatitis B has become a worldwide disease that seriously threatens human health, and it is also the most widespread and most serious infectious disease in China. At present, there are 350 million chronic hepatitis B virus (HBV) carriers in the world, and 121 million HBV carriers in my country, accounting for 9.75% of the national population. There are about 30 million chronic hepatitis B patients, and the annual death toll reaches 300,000. Moreover, there are about 500,000 new cases of hepatitis B in my country every year, accounting for about 1 / 4 of the number of infectious diseases in the country. At present, the annual cost for the prevention and treatment of liver diseases in the whole country is as high as more than 100 billion ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/06C07K1/36A61K39/42A61P1/16A61P31/12
Inventor 许贤豪
Owner 许贤豪