重组金葡萄球菌肠毒素B纯化工艺研究  

Purification of the recombinant Staphylococcal enterotoxin B

作　　者：邓小红[1] 郑玉玲[2] 曾政 姜永强[2] 

机构地区：[1]重庆工商大学药物化学与化学生物学研究中心,重庆400067 [2]军事医学科学院微生物流行病研究所,北京100071 [3]重庆市动物卫生监督总站

出　　处：《中国人兽共患病学报》2007年第1期27-30,52,共5页Chinese Journal of Zoonoses

基　　金：国家自然科学基金资助项目(No.30171091);北京市自然科学基金资助项目(No.7022031)

摘　　要：目的建立无降解、高纯度的重组金葡萄球菌肠毒素B(Staphylococcal enterotoxin B,SEB)的纯化工艺,并检测活性以考察纯化工艺对蛋白的影响。方法用PBV220表达载体诱导表达出重组SEB后,采用(NH4)2SO4粗纯—疏水层析—离子交换色谱的组合纯化法纯化该蛋白。纯化后的样品经Western-blot鉴定及动物致死实验检测活性。结果通过该纯化工艺得到了纯度高(纯度>99%),稳定性高的重组SEB,其活性与天然SEB蛋白无明显差异,保持了高活性。结论本实验建立了无降解、高纯度、高活性的重组SEB的纯化工艺,为进一步研究SEB的临床应用打下了基础。To set up an useful chromatography technique to obtain non-degraded recombinant Staphylococcal enterotoxin B （rSEB） with high purity and high biological activity. The recombinant SEB was expressed using prokaryotic expression vector PBV220 and then purified by a series of processes. The whole chromatography methods has three steps including ammonium sulfate precipitation , hydrophobic interaction chromatography, and SP-HP strong cation-exchange chromatography. The purified rSEB were identified by Western-blotting and its biological activity was evaluated by mouse lethality assay. As a result, non-degraded rSEB with high purity （purity〉99 %） was obtained, and the lethality assay in mice demonstrated that toxicity Of rSEB was the same as that of the natural SEB. It concludes that a good chromatography technique was set up which can obtained rSEB with high biological activity, high purity and high stability. This technique would provide a basis for further studies on the clinical application of SEB.

关 键 词：重组SEB 纯化工艺 生物活性 

分 类 号：R996.1[医药卫生—毒理学]