3种免疫学方法对肺炎球菌结合物及其中间品的多糖抗原性研究

Study on the antigenicity of Streptococcus pneumonia polysaccharide, its derivatives and conjugates by three immunological assays

目的：对肺炎球菌结合物及其制备过程中间品进行多种方法抗原性监控，以期对肺炎球菌结合物制备过程中多糖抗原性进行差异分析，进而为制备工艺参数的选择提供重要参考指标。方法用已建立的免疫双扩散法、抑制ELISA、速率比浊法对23 F型肺炎球菌结合物制备过程中间品和结合物进行抗原性分析；对不同方法结果比较、综合分析23 F型肺炎球菌结合物制备过程中的多糖抗原性变化。结果建立分析抗原性变化的抑制ELISA和速率比浊法；3种方法均可用于肺炎球菌结合物制备过程中的多糖抗原性分析；其中抑制ELISA可监控制备过程中抗原性的微小改变并可量化。结论用多种方法对23 F型肺炎球菌结合物制备过程中的多糖抗原性进行分析，为结合物制备过程中多糖抗原性的监控提供了多层面的有效方法，并为抗原性监控提供可量化的方法。

Objective To monitor and analyze the antigenicity of Streptococcus pneumonia polysaccharide, its derivatives and conjugates by three immunological assays .Methods Inhibition ELISA and rate nephelometry（RN） were established for this study.Antigenicity of serotype 23F pneumococcal conjugates and their intermediates were analyzed by double immunodiffusion assay , inhibition ELISA and RN .The results derived from three assays were comparatively analyzed to evaluate the changes of antigenicity during the preparing process of serotype 23F conjugate.Results Double immunodiffusion assay, inhibition ELISA and RN were all applicable to antigenicity analysis during the process of conjugate preparation .Inhibition ELISA could quantitatively detect a slight difference of polysaccharide antigenicity during the preparing process . Conclusion The antigenicity of polysaccharide during the preparing process of pneumococcal conjugates could be analytically monitored by using three immunological assays .This study provided evidence for successfully using immunological assays as the quality control means during the preparing process of pneumococcal conjugates .