Preparation and identification of a polyclonal antibody against Pgp3 of Chlamydia trachomatis
ZHOU Luqi, YANG Jia, LI Yang, ZHU Shanli.
Department of Microbiology and Immunology, Institute of MolecularViruses and Immunology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035,China
ZHOU Luqi,YANG Jia,LI Yang, et al. Preparation and identification of a polyclonal antibody against Pgp3 of Chlamydia trachomatis[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2024, 54(3): 184-189.
Abstract：Objective: To prepare a rabbit polyclonal antibody against Pgp3 protein of Chlamydia trachomatis (Ct) and determine its biological function in vitro. Methods: Recombinant protein Pgp3 was obtained by prokaryotic expression system and purified by nickel ion affinity chromatography. After renaturing in Phosphate buffered solution, the renatured Pgp3 protein was injected subcutaneously into Japanese-white-rabbits at multiple sites to generate polyclonal antibody. The titer and specificity of the polyclonal antibody was assessed by ELISA and Western blot, respectively. The specific recognition ability of the polyclonal antibody to natural Pgp3 protein was tested by indirect immunofluorescence assay (IFA) with Ct-infected HeLa cell model and Western blot under non-denaturing conditions. Results: The Pgp3 protein was successfully obtained and mainly expressed in the form of inclusion body. After immunization, the rabbits produced a high level of polyclonal antibody with a titer reached 1:512 000. Western blot and IFA showed that the rabbit polyclonal antibody against Pgp3 specifically recognized both the recombinant and native Pgp3 protein. Conclusion: The study successfully generated a high level of polyclonal antibody against Pgp3, which exhibited specific recognition abilities for both recombinant and native Pgp3 protein. These results provide a foundation of further research on the biological function of Pgp3 and the development of Ct immunotherapy agents.
