甲型肝炎病毒复合层析介质Capto Core 400的纯化效果  

Purification effect of composite chromatography medium Capto Core 400 on hepatitis A virus

作　　者：李春雷 岳立广 夏青娟 王艺博 王昕雷 徐艳玲 LI Chunlei;YUE Liguang;XIA Qingjuan;WANG Yibo;WANG Xinlei;XU Yanling(The Second Department of Vaccine in Changchun Institute of Biological Products Co.,Ltd.,Changchun 130012,Jilin Province,China)

机构地区：[1]长春生物制品研究所有限责任公司疫苗二室,吉林长春130012

出　　处：《中国生物制品学杂志》2023年第4期452-457,共6页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划项目(20190404001YY)。

摘　　要：目的 探索新型复合层析介质Capto Core 400层析填料纯化甲型肝炎病毒(hepatitis A virus,HAV)的效果。方法用Capto Core 400层析介质纯化人胚肺二倍体细胞(2BS株)培养收获的HAV粗纯液,经流速(60、150、300、450 cm/h)、上样缓冲液pH(6.6、7.0、7.4)、上样缓冲液盐离子浓度(0、0.15、0.3、0.45 mol/L)、载量[0.5、1、2、4柱体积(column volume,CV)]筛选最适纯化HAV的条件,并与传统Sepharose 4FF层析介质纯化效果进行比较。结果 初步确定Capto Core 400纯化工艺条件为流速150 cm/h、上样缓冲液pH 7.0~7.4、盐离子浓度0.15 mol/L、上样量2 CV时,纯化效果最好,病毒回收率平均为92.61%,蛋白去除率平均为84.25%。Sepharose 4FF层析介质纯化HAV抗原回收率平均为76.03%,蛋白去除率平均为73.21%,Capto Core 400层析介质纯化效果优于Sepharose 4FF层析介质。结论Capto Core 400复合层析介质纯化HAV回收率较高,操作相对简便,适合纯化HAV。Objective To explore the purification effect of hepatitis A virus(HAV)with a new type of composite medium Capto Core 400 chromatography packing.MethodsHAV crude liquid cultured with human embryonic lung diploid cells(2BS strain)was purified using Capto Core 400 chromatography media.The optimum conditions for purification of HAV were determined from flow rates(60,150,300 and 450 cm/h),pH of loading buffer(6.6,7.0 and 7.4),salt ion concentration of loading buffer(0,0.15,0.3 and 0.45 mol/L)and capacity[0.5,1,2 and 4 column volume(CV)],respectively.The purification effect was compared with that of traditional Sepharose 4FF chromatography media.Results The optimum purification conditions of Capto Core 400 were preliminarily determined as follows:flow rate of 150 cm/h,loading buffer pH of 7.0 ~ 7.4,salt ion concentration of 0.15 mol/L,loading volume of 2 CV.The average recovery rate of virus was 92.61%,and the average removal rate of protein was 84.25%.The average recovery rate of HAV antigen purified by Sepharose 4FF chromatography media was 76.03% and the average protein removal rate was 73.21%.The purification effect of Capto Core 400 chromatography media was better than that of Sepharose 4FF chromatography media.ConclusionThe Capto Core 400 composite chromatography media had a high recovery rate for HAV purification and a relatively simple operation,which was suitable for HAV purification.

关 键 词：甲型肝炎病毒 Capto Core 400 纯化 

分 类 号：R373.21[医药卫生—病原生物学]