Expression, purification of Staphylococcus aureus Efb protein and preparation of its functional antibody
- VernacularTitle:金黄色葡萄球菌Efb蛋白的表达、纯化及功能抗体的制备
- Author:
Xin, ZHANG
;
Ya-Ping, GAO
;
Jie, DONG
;
Yu, LIU
;
Na, WU
;
Ning-Sheng, SHAO
;
Guang, YANG
- Publication Type:Journal Article
- Keywords:
aureus,Staphylococcus;
Efb;
complement inhibition;
antibody
- From:Bulletin of The Academy of Military Medical Sciences
2009;33(6):531-534
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone and express the Staphylococcus aureus Efb(extracellular fibrinogen-binding protein) protein in Escherichia coli, to purify the expression product and prepare its functional antibody and to detect the functions of Efb protein for further studies on S.aureus infection.Methods: Efb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template and cloned into the recombinant expression vectors pET28a. E.coli BL21(DE3) with the plasmid was induced with IPTG for protein production. The protein was purified by Ni~(2+) affinity chromatography. The function of Efb protein was determined by complement activity assay and inhibition ELISA.The polyclonal antibodies were prepared by immunizing the animals. Results: The purified recombinant Efb was obtained, which could inhibit the CH50 and AH50 effectively. The functional poly-antibodies of Efb were prepared.Conclusion:Efb could inhibit the classical pathway and alternative pathway of complement activation, and the antibodies against to Efb could block the inhibition of the classical pathway of complement activation induced by Efb.
