Preparation and identification of polyclonal antibody against methyl-accepting chemotaxis signal transduction protein of Helicobacter hepaticus / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To prepare the polyclonal antibody against methyl-accepting chemotaxis signal transduction protein (MCP) of Helicobacter hepaticus (H.hepaticus).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>The fusion protein was successfully expressed, and the titer of the antibody reached 132 000. Western blotting indicated that the antibody could specifically bind to CSPs and His-rhMCP.</p><p><b>CONCLUSION</b>The antibody with a high titer and specificity was prepared to facilitate further study of the pathogenicity and epidemiology of H.hepaticus in human.</p>