免疫磁珠法分离preS1结合蛋白

Isolation of Hepatitis B Virus PreS1 Binding Protein by Immunomagnetic Beads

目的:用免疫磁珠分离人肝癌细胞(HepG2)细胞膜上与乙型肝炎病毒(HBV)preS1肽段结合的蛋白,建立快速分离膜受体蛋白的方法。方法:提取HepG2细胞膜蛋白,以生物素标记的preS1肽段为钓饵,与提取的膜蛋白4℃孵育过夜,加入链霉亲和素标记的磁珠,洗涤磁珠后收集结合蛋白,通过SDS-PAGE电泳分离结合蛋白。结果:用磁珠的方法所收集到的结合蛋白,通过SDS-PAGE电泳分离出了多条条带,比较明显的45～97 ku有9条,>97 ku处有5条,<20 ku处有2条。结论:通过免疫磁珠分离法可有效的获得HepG2细胞膜上preS1肽段的结合蛋白。

Objective： Using immunomagnetic beads method to separate the receptor- binding protein of HepG2 membrane that binding hepatitis B virus preS1 region, and to establish a fast method for separating membrane receptor proteins. Methods： Membrane protein was extracted from HepG2 cells, then combined with preS1 peptide which labeled with biotin, incubated overnight at 4 ℃ with the extraction of membrane proteins. After that, streptavidin-conjugated beads were added. After washing the beads and collection of binding proteins, proteins were separated by SDS-PAGE electrophoresis. Results： Binding proteins were collected by immunomagnetic beads and multiple stripes were separated by SDS-PAGE electrophoresis. There were nine stripes between 45 ku - 97 ku obviously, and five stripes in area lager than 97 ku, two stripes in area smaller than 20 ku. Conclusions： The preS1 peptide binding protein of HepG2 membrane can be effectively obtained by using immunomagnetic beads separation method .