摘要
旨在应用体积排阻色谱法测定口蹄疫灭活疫苗中的146S抗原含量。使用TSKgel G4000SWXL(7.8 mm×30 cm)色谱柱,以pH 7.2的缓冲盐体系作为流动相,流速为0.6 mL/min,进样量为100μL,检测波长为259 nm。以口蹄疫病毒(O型)灭活146S抗原纯化样品建立标准曲线;使用灭活抗原液配制3份口蹄疫灭活疫苗,并进行精密度、重复性、特异性、耐受性验证;应用该方法快速测定16批疫苗的146S含量。结果表明,抗原浓度在0.56–67.42μg/mL范围内,其峰面积与浓度的线性关系良好(R^2=0.996,n=10),3份疫苗146S抗原测定回收率分别为93.6%(RSD=2.7%,n=3)、102.3%(RSD=2.6%,n=3)、95.5%(RSD=5.1%,n=3),方法重复性好、准确性强(RSD=0.5%,n=6),且操作简便、高效,对16批疫苗的测定结果较理想。应用该方法有望快速高效地检测口蹄疫灭活疫苗的146S抗原含量,为疫苗的质量控制提供有力支持。
The aim of this study is to quantify the 146 S antigen in foot-and-mouth disease virus（FMDV） inactivated vaccine by size-exclusion chromatography（SEC）. The analysis was performed on a TSKgelG4000 SWXL column（7.8 mm×30 cm）, with a pH 7.2 buffer salt system as the mobile phase. The flow rate was 0.6 mL/min, the injection volume was 100 μL and the detection wavelength was 259 nm. The calibration curve was established by using purified inactivated FMDV（serotype O） 146S antigen; 3 batches of vaccine formulated by inactivated antigen solution were tested to verify the accuracy, reproducibility, specificity and tolerability of the method. At last 16 batches of vaccine were determined by the SEC method. Results showed a good linearity between peak area and concentration of 146S antigen in the range between 0.56 and 67.42 μg/mL（R^2=0.996, n=10）, and the average recovery rate of 146S antigen in the 3 batches of vaccine formulated in lab were 93.6%（RSD=2.7%, n=3）, 102.3%（RSD=2.6%, n=3）, and 95.5%（RSD=5.1%, n=3）. The method was proved accurate and reliable with good reproducibility（RSD=0.5%, n=6）, and applied to determine 16 batches of the commercial FMDV vaccine. According to the above results, the SEC method is high effective for 146 S antigen quantify in the inactivated FMDV vaccine and would provide strong support for the vaccine quality control.
作者
徐嫄
邹兴启
李翠
朱元源
何天慈
杨延丽
林旋
宋艳民
郑金来
张松平
赵启祖
Yuan Xu1, Xingqi Zou1, Cui Li1, Yuanyuan Zhu1, Tianci He1, Yanli Yang2, Xuan Lin2, Yanmin Song2, Jinlai Zheng3, Songping Zhang2, and Qizu Zhao1(1 China Institute of Veterinary Drug Control, Beijing 100081, China; 2 National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China; 3 Beijing Biaochizehui Biological Technology Co. Ltd., Beijing 102600, Chin)
出处
《生物工程学报》
CAS
CSCD
北大核心
2018年第5期676-684,共9页
Chinese Journal of Biotechnology
基金
国家重点研发计划(No.2016YFD0501500)
公益性行业(农业)科研专项经费资助(No.201303046)资助~~
关键词
体积排阻色谱
口蹄疫
灭活疫苗
146S抗原
size-exclusion chromatography
foot-and-mouth disease
inactivated vaccine
146S antigen
