Purification and immunological characteristics of monoclonal antibody 2H4 against Chlamydia trachomatis pORF5 plasmid protein
10.3760/cma.j.issn.0254-5101.2011.11.019
- VernacularTitle:沙眼衣原体pORF5质粒蛋白单克隆抗体2H4的纯化及其免疫学特性研究
- Author:
Zhongyu LI
;
Yimou WU
;
Qiulin HUANG
;
Shengmei SU
;
Zhou ZHOU
;
Chaoqun CHEN
;
Hui ZHOU
;
Guangming ZHONG
- Publication Type:Journal Article
- Keywords:
Chlamydia trachomatis;
pORF5 plasmid protein;
Monoclonal antibody;
Immunological characteristics
- From:
Chinese Journal of Microbiology and Immunology
2011;31(11):1041-1045
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo purify and characterize the monoclonal antibody (McAb) against Chlamydia trachomatis pORF5 plasmid protein.Methods The hybridoma cells stably secreting specific McAb against pORF5 were cultured in a large scale,and protein G purification by affinity chromatography was used to purify 2H4 McAb.ELISA was used to determine the antibody titer,and identify McAb isotype.Immunofluorescence assay (IFA) and Western blot were performed to detect McAb specificity.Results The purity of 2H4 antibody was 93％,the titer reached 1:1024,and 2H4 McAb was identified to belong to IgG2a isotype,2H4 McAb reacted strongly with the GST-pORF5 fusion protein and endogenous pORF5 protein expressed by Chlamydia trachomatis serovar A,D,L2,Chlamydia muridarum ( MoPn ),Chlamydia psittaci 6BC,but not other chlamydial plasmid proteins and Chlamydia pneumoniae(Cpn) AR39 strain.Conclusion2H4 McAb against pORF5 protein was successfully purified with a high titer and specificity which lay a foundation for further study on pORF5 protein structure and function.
