关键词:

沙眼衣原体, 主要外膜蛋白, 多肽, 多克隆抗体

Abstract:

Objective The major outer membrane protein（MOMP）of Chlamydia trachomatis（Ct）is essential for Ct structural stability and infection of the host cell. MOMP polyclonal antibody was created using the synthetic peptide to further investigate the function and mechanism of MOMP protein. Methods Ct MOMP antigenic epitope was investigated. MOMP⁃specific peptide was designed and synthesized based on the conserved peptides of 15 different Ct serotype MOMPs，and New Zealand white rabbits were immunized after coupling with hemocyanin. MOMP⁃specific peptide was designed and synthesized based on the conserved peptides of 15 different Ct serotype MOMPs，and New Zealand white rabbits were immunized after coupling with hemocyanin.Immunoaffinity chroma⁃ tography was used to separate the immune serum. SDS⁃PAGE was used to determine the purity of the IgG antibody， and ELISA was used to determine the titer of the MOMP antibody. Furthermore，Western blot and immunofluores⁃ cence staining were used to determine the specificity of the MOMP antibody. Results The purity of IgG antibody was 89.6%，and the titer of MOMP antibody was 1∶12800. The results of Western blot showed that the antibody specifically bound to 8 serotypes of Ct protein at 40kDa，and indirect immunofluorescence staining showed that the antibody could specifically combined with Ct MOMP. In addition，the immunofluorescence results showed that the MOMP antibody and the imported commercial antibody were obviously co ⁃localized in the host cells ，and the fluorescence intensities of the two antibodies were comparable. Conclusions This study successfully prepared arabbit polyclonal antibody with high titer and good specificity against Ct MOMP ，providing a useful tool for future research on the biological function of MOMP.

Key words:

Chlamydia trachomatis, major outer membrane protein, peptide, polyclonal antibody