Preparation and identification of monoclonal antibodies against pgp3 of Chlamydia trachomatis
10.3969/j.issn.1000-484X.2015.05.017
- VernacularTitle:沙眼衣原体质粒蛋白pgp3单克隆抗体制备及鉴定
- Author:
Xiaohui JIA
;
Fang ZHOU
;
Xia ZHAO
;
Ting LI
;
Tianjun JIA
- Publication Type:Journal Article
- Keywords:
Chlamydia trachomatis;
Secreted protein;
Plasmid protein;
Monoclonal antibody
- From:
Chinese Journal of Immunology
2015;(5):659-662
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express secreted protein-pgp3 of Chlamydia trachomatis(Ct)plasmid,produce monoclonal antibodies (mAbs)and identify their basic biological characteristics.Methods: Construction pGEX-6p2-pgp3 prokaryotic expression vector,then expressed GST-pgp3 fusion protein in E.coli as antigen used to immune BALB/c mice, spleen cells were fused with SP2/0 mouse myeloma cells.The hybridoma cell lines of screening mAbs were secreted by ELISA,and mAbs specificity,type,class and titer were identified.Results:GST-pgp3 fusion protein was successful expressed,5 strains stable hybridoma cell lines that secreted mAbs were screened out,including 4 strains secreted anti-pgp3 mAbs(P1B3,P2A1,P2B6,P2C2),mAbs type were IgG1/κ,the other strain secretion anti-GST mAbs(P3B4),mAbs type was IgG2b/κ.The titer of P1B3,P2A1,P2B6,P2C2,P3B4 were 1∶6 400,1∶3 200,1∶12 800,1∶6 400 and 1∶6 400 respectively.Conclusion:Successful prepared anti-pgp3 mAbs,and lay a foundation for further study the function of Ct plasmid protein pgp3 and the establishment of Ct detection method.
