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中华仓鼠卵巢细胞(CHO)表达的重组乙肝病毒表面抗原(HBs Ag)为糖基化蛋白。上游主要使用细胞贴壁培养工艺,下游主要使用疏水、离子交换、凝胶过滤层析。但由于培养后期,部分细胞脱落死亡,细胞中杂蛋白释放到上清液中,在纯化中影响离子介质载量、抗原收率和介质寿命,普通清洗方法不能有效恢复。本研究分别用70%乙醇+0.5M氢氧化钠、1M、2M氢氧化钠[1],与原有清洗方法0.5M氢氧化钠和新介质对比,对到达使用寿命的旧离子介质进行清洗,后相同条件进行层析。最终发现,2.0M氢氧化钠清洗后的介质,载量和收率均恢复,且原液质量符合标准要求。这不仅提高了收率、保证了原液质量,而且延长了介质寿命。
The recombinant hepatitis B virus surface antigen (HBs Ag) expressed by Chinese hamster ovary cells (CHO) is a glycosylated protein. Upstream, the main use of cell adherent culture process, the downstream use of hydrophobic, ion exchange, gel filtration chromatography. However, due to late cell culture, part of the cells shed and died, and the extracellular proteins in the cells are released into the supernatant, influencing the ion medium loading, the yield of antigen and the life span of the medium in purification, and the ordinary washing method can not be effectively recovered. In this study, 70% ethanol + 0.5M sodium hydroxide, 1M, 2M sodium hydroxide [1], compared with the original cleaning method 0.5M sodium hydroxide and new media, to reach the service life of the old ion medium cleaning, After the same conditions for chromatography. Finally found, 2.0M sodium hydroxide cleaning media, load and yield were restored, and the quality of the stock solution meets the standard requirements. This not only improves the yield, ensures the quality of the liquid, but also prolongs the life of the medium.