亲和层析法纯化霍乱毒素及其B亚单位

利用GM1偶联活化的Sephadex G50或扰CT IgG偶联活化的Sepharose 4B,我们已成功地建立了两种亲和层析纯化CT和CT-B的方法,获得了纯化产品。受体亲和柱对CT-B和CT的分离效率分别为71.6％或48.3％,抗体亲和柱分别为50％或51.7％。纯化产物经SDSPAGE、PAGE分析表明均达到亲和层析电泳纯级。GM1-ELISA、CHO细胞测毒加间接免疫荧光检测、家兔肠段结扎试验和家兔免疫保护试验结果提示纯化产物具有良好的生物学活性。这种有效而简单的方法可推广应用于CT或CT相关毒素的纯化。

Purification of CT and CT-B by Affinity Chromatography Methods

We developed two kinds of affinity chromatography methods for isolating CT and CT-B with GM1 coupling to activated Sephadex G_(50) or anti-CT IgG coupling to activated Sepharose 4B respectively. The isolating effect of receptor affinity chromatogragy for CT-B or CT are 71.6% or 48.3% respectively. IgG affinity column for CT-B or CT are 50% or 51.7% respectively. The results of SDS-PAGE and PAGE analysis showed that the purified CT-B and CT reached to the electrophoresis purification degree of affinity chromatography. By GM1-ELISA, CHO cytotoxic test, cellular indirect immunofluorescenee, rabbit ileal loop test and rabbit immunoprotective test, the purified CT-B and CT have been proved that the both have good bioactivities. The simpler effective methods may be applied to the purification of CT or CT related enterotoxins.