狂犬疫苗糖蛋白ELISA定量分析方法的建立及验证  

Establishment and Verification of ELISA Quantitative Analysis Method for Rabies Vaccine Glycoprotein

作　　者：叶珂 张志高 何春辉 方浩霖 林冠峰 卞论 许四宏 吴英松 YE Ke;ZHANG Zhigao;HE Chunhui;FANG Haolin;LIN Guanfeng;BIAN Lun;XU Sihong;WU Yingsong(Key Laboratory of Antibody Engineering of Guangdong Higher Education Institutes,School of Laboratory Medicine and Biotechnology,Southern Medical University,Guangzhou 51515,China;Guangzhou Nuocheng Biological Products Co.,Ltd,Guangzhou 511495,China;National Institutes for Food and Drug Control,Beijing 100050,China)

机构地区：[1]南方医科大学检验与生物技术学院广东省高等学校抗体工程重点实验室,广州510515 [2]中国食品药品检定研究院,广州511495 [3]广州诺诚生物制品有限公司,北京100050

出　　处：《病毒学报》2022年第1期8-13,共6页Chinese Journal of Virology

基　　金：广州市协同创新重大专项(项目号:201400000004-1),题目:重大传染病创新检测试剂的研制和应用研究。

摘　　要：为狂犬疫苗生产质量控制建立简便、高效、精准的糖蛋白体外效力分析方法,研制适用于狂犬疫苗生产质量控制的狂犬病毒糖蛋白酶联免疫吸附(ELISA)定量分析试剂。采用双抗体夹心法建立狂犬病毒糖蛋白ELISA定量分析体系,优化反应各项参数后考核试剂性能。线性范围为1~180 mIU/mL,灵敏度为0.4497 mIU/mL,分析内变异系数为5.8%~8.2%,分析间变异系数为7.2%~14.4%。用自制试剂与National Institutes of Health(NIH)动物法平行检测10份疫苗相关样品,两种方法测定结果的相对系数r^(2)为0.8943,相关性良好。自制ELISA试剂各项性能指标良好,且与NIH动物法检测结果相关性良好,有望推广用于狂犬疫苗生产质量控制。To provide a simple,efficient and accurate in vitro potency analysis method for glycoprotein production quality control for rabies vaccine production,and to develop a quantitative analysis reagent for rabies virus glycoprotein linked immunosorbent assay(ELISA)suitable for rabies vaccine production quality control.The double antibody sandwich method was used to establish the rabies virus glycoprotein ELISA quantitative analysis system.After the optimization of each reaction parameter,the performance of the reagent was evaluated.The linear range was 1 mIU/mL to 180 mIU/mL,the sensitivity was 0.4497 mIU/mL,the intraanalysis coefficient of variation was 5.8%to 8.2%,and the inter-analysis coefficient of variation was 7.2%to14.4%.10 vaccine-related samples were tested in parallel with the National Institutes of Health(NIH)animal method using self-made reagents.The relative coefficient r^(2)of the results of the two methods was 0.8492,and the correlation was good.The performance indicators of the self-made ELISA reagents are good,and they have a good correlation with the NIH animal test results.It is expected to be popularized for the quality control of rabies vaccine production.

关 键 词：狂犬疫苗 糖蛋白 质量控制 双抗体夹心酶联免疫吸附试验(ELISA) 

分 类 号：R373.9[医药卫生—病原生物学]