人二倍体细胞狂犬疫苗两种纯化方式比较分析

Comparative analysis of two purification methods of human diploid cell rabies vaccine

目的评价人二倍体细胞狂犬疫苗两种纯化方式的效果。方法比较分析分子筛层析柱法和密度梯度离心法两种纯化方式得到的狂犬病毒纯化液的抗原回收率、蛋白含量和主要杂质去除率来判断这两种方式的纯化效果。结果分子筛层析柱法得到的纯化液抗原回收率均值为69.8%,蛋白含量均值为179.27μg/mL,牛血清白蛋白去除率高于99.98%;密度梯度离心法得到的纯化液抗原回收率均值为76.7%,蛋白含量均值为144.82μg/mL,牛血清白蛋白去除率高于99.98%。结论两种方式得到的狂犬病毒纯化液都符合要求,但是密度梯度离心法的总蛋白含量低、抗原回收率高,相较于分子筛层析柱法纯化效果更佳。

Objective To evaluate the efficacy of two purification methods of human diploid cell rabies vaccine.Methods By analyzing and comparing the protein content,antigen recovery rate and main impurity removal rate of the virus purification solution obtained by density gradient centrifugation and chromatography column purification,the purification efficiency of the two purification methods was evaluated.Results Based on the method of density gradient centrifugation purification,the average antigen recovery rate and protein content were 69.8% and 179.27μg/mL,respectively,and the removal rate of bovine serum albumin was higher than 99.98%.Based on the method of chromatography column purification,the average antigen recovery rate and protein content were 76.7% and 144.82μg/mL,respectively,and the removal rate of bovine serum albumin(BSA)was higher than 99.98%.Conclusion The antigen content,protein content and BSA residue of the purified solution obtained by the two methods all meet the requirements of the pharmacopoeia.However,the density gradient centrifugation purification method has a lower total protein content and a higher antigen recovery rate,and has a better effect than the chromatography column purification method.