铝吸附EV71-CA16二价手足口病灭活疫苗CA16抗原解离方法研究  

A method for antigen dissociation of CA16 antigen in EV71-CA16 bivalent inactivated vaccine containing aluminium adjuvant

作　　者：张浩然 曹明翔 刘鹏 徐华 陈璠瑷 杨二霞 ZHANG Haoran;CAO Mingxiang;LIU Peng;XU Hua;CHEN Fanai;YANG Erxia(Research and Development Department,Aimei Action Biopharmaceutical Co.,Ltd.,Taizhou Jiangsu,225300,China)

机构地区：[1]艾美行动生物制药有限公司研发部,江苏泰州225300

出　　处：《质量安全与检验检测》2024年第6期107-111,共5页QUALITY SAFETY INSPECTION AND TESTING

摘　　要：为建立铝吸附EV71-CA16二价手足口病灭活疫苗CA16抗原解吸附的方法,用于CA16抗原含量的检测。本文采用单一变量的方法,分别固定柠檬酸钠、10%曲拉通与20%二乙醇胺的比例以确定最佳解离液配方,在此基础上通过不同孵育温度和时间处理疫苗成品,ELISA法检测解离后的CA16抗原含量,计算抗原回收率,确定最佳解离液配方及解离条件,并考察其重复性。结果显示,解离液最佳方法为20%二乙醇胺1.25 mL、10%曲拉通50μL、柠檬酸钠浓度为14%,由0.01M PBS定容至10 mL,室温(18~25℃)孵育0.5 h解离疫苗成品,经解离后的CA16的抗原含量达1800 U/mL,回收率在90%以上,线性相关系数R2值大于0.97,重复试验的变异系数均小于5%。结果表明,该解离方法回收率高,重复性好,检测结果准确。To develop a method for CA16 antigen dissociation and content determination of EV71-CA16 bivalent inactivated vaccine containing aluminiuma djuvant for the detection of CA16 antigen content.The ratio of sodium citrate,10%tramadol and 20%diethanolamine was fixed by single variable method to determine the best formula of dissociation solution,on this basis,the vaccine was treated with different incubation temperature and time,the content of CA16 antigen after dissociation was detected by ELISA,the antigen recovery rate was calculated,the best dissociation solution formula and dissociation conditions were determined,and its repeatability was investigated.The results showed that the best formula of dissociation solution is 20%diethanolamine 1.25 mL,10%tralatone 50μL,the concentration of sodium citrate is 14%,and the final volume is fixed to 10 mL with 0.01 M PBS,the best dissociation condition is incubation at room temperature(18-25℃)for 0.5 h.After dissociation,the antigen content of CA16 reached 1800 U/mL,the recovery rate was more than 90%,the linear correlation coefficient R2 was more than 0.97,and the coefficient of variation of repeated tests was less than 5%.The results indicate that the dissociation method has high recovery,good repeatability and accurate detection results.

关 键 词：EV71-CA16二价手足口病灭活疫苗 柯萨奇病毒A组16型(CA16) 解离液 铝佐剂 ELISA 

分 类 号：R392-33[医药卫生—免疫学]