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
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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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