TY - JOUR
T1 - Purification and immunological characteristics of monoclonal antibody 2H4 against Chlamydia trachomatis pORF5 plasmid protein
AU - Li, Zhong Yu
AU - Wu, Yi Mou
AU - Huang, Qiu Lin
AU - Su, Sheng Mei
AU - Zhou, Zhou
AU - Chen, Chao Qun
AU - Zhou, Hui
AU - Zhong, Guang Ming
PY - 2011/11/1
Y1 - 2011/11/1
N2 - Objective: To 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. ELTSA 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 TgG2a 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. Conclusion: 2H4 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.
AB - Objective: To 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. ELTSA 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 TgG2a 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. Conclusion: 2H4 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.
KW - Chlamydia trachomatis
KW - Immunological characteristics
KW - Monoclonal antibody
KW - PORF5 plasmid protein
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U2 - 10.3760/cma.j.issn.0254-5101.2011.11.019
DO - 10.3760/cma.j.issn.0254-5101.2011.11.019
M3 - Article
AN - SCOPUS:84864750786
SN - 0254-5101
VL - 31
SP - 1041
EP - 1045
JO - Chinese Journal of Microbiology and Immunology (China)
JF - Chinese Journal of Microbiology and Immunology (China)
IS - 11
ER -