摘要
我们成功地建立了层析等电聚焦的三步程序,分离提纯葡萄球菌D型肠毒素。首先用CG-50树脂吸附由细菌培养液中获得粗毒素,然后在PBE94柱上进行层析等电聚焦,最后经过Sephacryls-200过滤。纯化的SED纯度约98%,回收率89%,分子量为28500,pI 7.6。Western blot,免疫双向琼脂扩散的鉴定试验表明,纯化SED与其相应抗血清是特异性反应。
A chromatofocusing procedure for the purification of Staphylococcal en-terotoxin D(SED) has been developed. The purification includes the removal of the toxic protein from the supernatant of culture of 1151m by batch absorption with CG-50 resin,chromatofocusing on PBE94 and gel permeation chromatography on Sephacryl S-200 respectively. The purity of SED using this procedure was 98%. The recovery of SED was 89% from chromatofocusing to Sephacryl S-200. Its molecular weight was determined to be 28 500 by SDS-PAGE and isoelectric point of 7.6 as determined by isoeloctric focusing. Western blot, Immunodiffusion testing revealed immunolo gical specific reaction of purified SED.
