摘要
目的纯化大肠杆菌表达的重组幽门螺杆菌粘附素。方法将表达HpaA蛋白的工程菌经高压均质机破菌获得包涵体,经洗涤、变性、复性,Q Sepharose High Performance阴离子交换层析和亲和层析分离纯化。采用SDS-PAGE和HPLC检测纯度,用Western blot检测其抗原性。结果纯化后HpaA蛋白纯度高达95％以上,具有良好的抗原性。结论建立了从包涵体中获得高纯度HpaA纯化工艺,为进一步的研究打下了基础。
Objective To establish an effective method for the purification of recombinant Helicobact-er pylori adhesin protein (rHpaA) expressed in E. coli. Methods The recombinant strain expressing HpaA protein was disrupted by Hyperbric homogeniser. The obtained inclusion bodies were washed, de-naturalized and re-naturalized, and the HpaA in them were purified by Q Sepharose High Performance negative ion exchange and affinity chromatography. The purity of HpaA was detected by SDS-PAGE and HPLC, and its anti-genicity by Western blot.Results The purified HpaA protein reached a purity of above 95% and showed good antigenicity. Conclusion An procedure for high purification of HpaA from inclusion body was developed.
出处
《中国生物制品学杂志》
CAS
CSCD
2004年第3期145-148,共4页
Chinese Journal of Biologicals
基金
国家"九五"重点科技攻关课题(96-901-01-54)
"八六三"生物与现代农业技术领域生物工程技术主题课题(2001AA21516).
