分子筛层析法分析重组乙肝表面抗原聚集物的形成

Analysis of recombinant hepatitis B surface antigen aggregate by size-exclusion chromatography

分子筛层析作为分析蛋白质颗粒聚集物的一种有力工具,被用于研究重组乙肝表面抗原聚集物的形成。已去除聚集物的表面抗原放置在不同的理化条件下或经过不同的纯化方法处理后,应用HPLC分析其聚集物的形成。为研究发酵过程中是否形成表面抗原聚集物,酵母细胞破碎后立即用Sepharose 4 FF层析柱分离为不同的组分,并分别进行HPLC分析。结果发现,在纯化过程和酵母发酵阶段都有表面抗原聚集物的产生。

Size-exclusion chromatography(SEC) is a powerful tool to analyze protein particle assembly.This approach was used to investigate the source of aggregation of recombinant hepatitis B surface antigen(HBsAg).Aggregate-free HBsAg was analyzed by HPLC after stored in different physico-chemical conditions or treated with different purification methods.To test whether aggregation takes place in vivo,crude yeast extract containing the expressed HBsAg was fractioned on a Sepharose 4 FF column just after cell disruption,and each fraction was analyzed by HPLC individually.As a result,The HBsAg aggregation takes place not only in purification process,but also in vivo.