[Expression of human anti-rabies Scds-Fv fusion protein in E.coli].
OBJECTIVE
To express the anti-rabies MBP-ScdsFv fusion protein in E.coli and identify it's activity assay.


METHODS
A full length of ScdsFv gene was Cloned into prokaryotic vector pMAL-p2x by recombinant DNA, Then the recombinant plasmid pMAL-ScdsFv was transformed to E.coli ER2566 for expression and induced by IPTG. The fusion protein was digested by Factor Xa, purified by Ni(+) and amylose resin affinity chromatography and Identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA.


RESULTS
Recombinant plasmid pMAL-ScdsFv was successfully constructed. The anti-rabies ScdsFv protein which was digested by Factor Xa and purified by Ni-NTA and amylose resin affinity chromatography had good binding activity and high stability.


CONCLUSIONS
The anti-rabies ScdsFv protein is of good binding activity and high stability, and it can be used for the immune therapy.
Cai
Kun
K
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China. ckreal@163.com
Wang
Hui
H
Shi
Jing
J
Bao
Shi-zhong
SZ
Hou
Xiao-jun
XJ
Yin
Jun
J
D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011820:Rabies virus / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
D011993:Recombinant Fusion Proteins