Preparation and quality control of four types of serotypes pneumococcal immune serum
Objective:To prepare type 6A,6B,19A,and 19F pneumococcal immune serum and examine their qualities to provide high-quality test reagents for the determination of polysaccharide content in pneumococcal vaccine.Methods:Type 6A,6B,19A and 19F pneumococcal immune serum was prepared by using type 6A,6B,19A and 19F pneumococcal strains as immunogenes,inoculated with white rabbit according to immunization procedures,and the prescribed immune serum was aliquoted certified and stored.Then the pneumococcal immune serum was detected by rate nephelometry and ELISA.The working concentration of pneumococcal immune serum was determined by external standard method,the specificity of immune serum was analyzed by cross reaction,and the preliminarily screened immune serum was absorbed and screened again by serum cross test.ELISA method was used to detect and compare the antibody titers of self-made pneumococcal immune serum with those produced by SSI in Denmark.Finally,the immune serum with cross-reaction with C polysaccharide was screened and absorbed.Results:Cross-reaction was detected in type 6A and 6B,19A and 19F serum,the cross-reaction was successfully removed after absorption,and the reaction curve was linear in the polysaccharide content range of 0.75～2.0 μg·mL-1(r>0.985).ELISA method was used to determine the antibody titer of the self-made pneumococcal immune serum and that produced by SSI in Denmark,and it was found that the self-made pneumococcal immune serum had a good antibody level.Type 19F serum had cross-reaction with C polysaccharide,while the response values of type 6A,6B and 19A serum had no significant change compared with the blank control.When C polysaccharide was added to type 19F serum for absorption,type 19F serum with an equal amount of diluent was prepared in parallel,and the response values before and after absorption were compared.The results were consistent,which proved that the effective components of 19F antibody would not be reduced by absorption and could be continued to be used according to the working dilution determined before absorption.Conclusion:The self-made pneumococcal immune serum of type 6A,type 6B,type 19A and type 19F had no cross reaction and the antibody level was good,which could be used for the determination of polysaccharide content of pneumococcal vaccine.
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