测定13价肺炎球菌结合疫苗多糖含量的速率比浊法的建立及验证  

Development and validation of rate nephelometry for determination of polysaccharide content in 13-valent pneumococcal conjugate vaccine

作　　者：韩菲[1] 郝倩[1] 张亭[1] 尹珊珊[1] 刘建凯[1] HAN Fei;HAO Qian;ZHANG Ting;YIN Shan-shan;LIU Jian-kai(Beijing Minhai Biotechnology Co.Ltd.,Beijing 102600,China)

机构地区：[1]北京民海生物科技有限公司,北京102600

出　　处：《中国生物制品学杂志》2020年第7期824-828,共5页Chinese Journal of Biologicals

摘　　要：目的建立速率比浊法测定13价肺炎球菌结合疫苗多糖含量,并进行验证。方法采用免疫化学分析系统(IMMAGE 800),用氢氧化钠解吸附处理样品,测定13价肺炎球菌结合疫苗多糖含量,并对建立的方法进行线性、精密性、准确性、专属性及溶液稳定性验证。结果该方法在本研究条件下,1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F、23F型肺炎球菌荚膜多糖浓度在0.5~3.0μg/mL范围内,线性良好,相关系数(r)均大于0.9850;各型多糖含量重复检测的相对标准差(RSD)均低于10%,中间精密性检测的RSD均低于15%;各型多糖含量检测的加标回收率在80%~120%之间;2、8、9N、10A、11A、12F、15B、17F、20、22F、33F型肺炎球菌精制多糖在曲线浓度范围内不影响13价肺炎球菌结合疫苗各型多糖含量的检测,且19A与19F之间、6A与6B之间不存在交叉反应;13价肺炎球菌结合疫苗稀释后4 h以内测定多糖含量结果稳定。结论速率比浊法测定13价肺炎球菌结合疫苗结果准确,精密性好,可作为该疫苗的质控方法。Objective To develop and validate a rate nephelometry for determination of polysaccharide content in 13-valent pneumococcal polysaccharide conjugate vaccine.Methods By using immunochemistry system(IMMAGE 800),13-valent pneumococcal polysaccharide conjugate vaccine was desorbed with sodium hydroxide and determined for polysaccharide content.The developed method was validated for linearity,precision,accuracy,specificity and solution stability.Results Under the condition in this study,the working curve showed good linearity at concentrations of 13 types(1,3,4,5,6 A,6 B,7 F,9 V,14,18 C,19 A,19 F,23 F)of pneumococcal capsular polysaccharide of 0.5~3.0μg/mL,with a correlation coefficient(r)of more than 0.9850.The relative standard deviations(RSDs)of determination results of polysaccharide of various types in repeat test were less than 10%.The spike recoveries of various types of polysaccharides were 80%~120%.The concentrations of purified polysaccharides of types 2,8,9 N,10 A,11 A,12 F,15 B,17 F,20,22 F and 33 F at the linear range on working curve showed no influence on the determination of polysaccharide content in 13-valent pneumococcal polysaccharide conjugate vaccine,while no cross reactions were observed between types 19 A and 19 F or 6 A and 6 B.The determination result of polysaccharide content was stable within4 h after dilution of 13-valent pneumococcal polysaccharide conjugate vaccine.Conclusion The developed rate nephelometry is accuracy and precise,which may be used for quality control of 13-valent pneumococcal polysaccharide conjugate vaccine.

关 键 词：速率比浊 13价肺炎球菌结合疫苗 多糖含量 方法学验证 

分 类 号：R917[医药卫生—药物分析学]