Purification effect of composite chromatography medium Capto Core 400 on hepatitis A virus / 中国生物制品学杂志

ABSTRACT

@#ObjectiveTo 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.