国产与进口琼脂糖凝胶过滤层析介质对Omicron株新型冠状病毒疫苗分离纯化效果的比较

Comparison of domestic and imported agarose gel filtration chromatography media for separation and purification of SARS⁃CoV⁃2 Omicron vaccine

目的探讨国产与进口琼脂糖凝胶过滤层析介质对Omicron株新型冠状病毒(SARS-CoV-2)灭活疫苗的分离纯化效果,为促进层析介质的国产替代提供数据支持。方法选取3批SARS-CoV-2 Omicron株灭活病毒浓缩液,分别使用国产与进口琼脂糖凝胶过滤层析介质在相同实验条件下进行纯化,收集病毒峰并检测收集组分中的抗原含量、蛋白质含量、Vero细胞DNA残留量、Vero细胞宿主蛋白残留量、牛血清白蛋白残留量等相关质量参数,并计算比活性和抗原回收率。结果在采用10%柱体积和30 cm/h线性流速对两种填料进行上样纯化之后,所得收集组分的平均抗原回收率分别为55.29%和53.31%,差异无统计学意义(t=1.44,P>0.05),比活性、Vero细胞DNA残留量及Vero细胞宿主蛋白残留量间差异也无统计学意义(t分别为-0.25、1.31和0.66,P均>0.05),仅牛血清白蛋白残留量差异有统计学意义(t=-2.73,P<0.05)。结论采用国产与进口琼脂糖凝胶过滤层析介质纯化SARS-CoV-2 Omicron株灭活病毒浓缩液,所得收集组分各项质量参数间除牛血清白蛋白残留量外均无显著差异,因此,在纯化Omicron株SARS-CoV-2灭活疫苗时,可考虑使用该国产层析介质替代进口层析介质。

Objective To explore the separation and purification efficiency of domestic and imported agarose gel filtration chromatography media for the inactivated SARS-CoV-2 Omicron vaccine,in order to provide data support for promoting the domestic substitution of chromatography media.Methods Three batches of concentrated SARS-CoV-2 Omicron strain inactivated virus were used in the experiments,and the purification was conducted using domestic and imported agarose gel filtration chromatography media under identical experimental conditions respectively.The virus peaks were collected,and relevant quality parameters,including antigen content,protein content,residual DNA of Vero cells,residual host proteins of Vero cells,and residual bovine serum albumin content in the collected components,were measured.The specific activity and antigen recovery rates were calculated.Results After loading and purification using a 10%column volume and a linear flow rate of 30 cm/h for both media,the average antigen recovery rates for the collected components were 55.29%and 53.31%,respectively,with no significant difference(t=1.44,P>0.05).Moreover,there was no significant difference in specific activity,residual DNA of Vero cells and residual host proteins of Vero cells(t=-0.25,1.31 and 0.66,respectively,each P>0.05).Only the residual bovine serum albumin content exhibited a significant difference(t=-2.73,P<0.05).Conclusion The domestic and imported agarose gel filtration chromatography media were used to purify the concentrated solution of SARS-CoV-2 Omicron strain inactivated virus,and the quality parameters of the collected components had no significant difference except for the residual bovine serum albumin content.Therefore,in the purification of the inactivated SARS-CoV-2 Omicron vaccine,the domestic chromatography media could be considered as a viable alternative to imported media.