[Preparation and identification of polyclonal antibody against methyl-accepting chemotaxis signal transduction protein of Helicobacter hepaticus].
OBJECTIVE
To prepare the polyclonal antibody against methyl-accepting chemotaxis signal transduction protein (MCP) of Helicobacter hepaticus (H.hepaticus).


METHODS
The recombinant plasmid pET22b+/MCP was transformed into E.coli BL2l(DE3) to express the fusion protein His-rhMCP under the induction of IPTG. The fusion protein was purified and the antibody was obtained by immunizing rabbits. The titer of the polyclonal antibody was tested by indirect ELISA, and the specificity of the antibody was identified based on Western blotting using the prepared cell surface proteins (CSPs) of the bacteria.


RESULTS
The fusion protein was successfully expressed, and the titer of the antibody reached 1:32 000. Western blotting indicated that the antibody could specifically bind to CSPs and His-rhMCP.


CONCLUSIONS
The antibody with a high titer and specificity was prepared to facilitate further study of the pathogenicity and epidemiology of H.hepaticus in human.
Department of Cardiovascular Thoracic Surgery, Nanfang Hospital, Sourthern Medical Universtity, Guangzhou 510515, China.E-mail: heshengping1982@163.com.
Shengping
S
He
Yahua
Y
Chen
Yingying
Y
Zhao
Jide
J
Wang
Yang
Y
Bai
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis* / Q000235:genetics; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000276:immunology; D044923:Helicobacter hepaticus / Q000276:immunology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D015398:Signal Transduction
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins