金黄色葡萄球菌B型肠毒素的培养产毒与柱层析纯化

Studies on the Production and Purification of Staphylococcal Enterotoxin B

本文采用玻璃纸覆盖琼脂平板法,在37℃培养金黄色葡萄球菌48h后,收集菌体,10000×g离心15min,上清液经聚乙二醇20000浓缩。浓缩液依次经羧甲基阳离子交换纤维素CM-32层析和葡聚糖凝胶SephadexG-75过滤纯化,获得电泳纯的金黄色葡萄球菌B型肠毒素。实验证明,利用该简化的二步柱层析分离得到的金黄色葡萄球菌B型肠毒素完全能满足血清学检实验和鉴定的要求,方法简便易行,一般实验室均可使用。

By the method of cellophane covered agar plate culture, production of staphylococcal enterotoxin B was studied. After incubation at 37℃ for 48h, cells were harvested and the cell suspension was centrifuged at 10000×g for 15min at 4℃. The supernatant was condensed by polyglycol 20000. Then condensation was purified to electrophoretic homogeneity of the staphylococcal enterotoxin B by ion-exchange chromatography on CM-cellulose and by gel filtration on Sephadex G-200. This method was simple and could be applicable for laboratory.