Purification and immunological characteristics of monoclonal antibody 2H4 against Chlamydia trachomatis pORF5 plasmid protein

Zhong Yu Li, Yi Mou Wu, Qiu Lin Huang, Sheng Mei Su, Zhou Zhou, Chao Qun Chen, Hui Zhou, Guangming Zhong

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1041-1045
Number of pages5
JournalChinese Journal of Microbiology and Immunology (China)
Volume31
Issue number11
DOIs
StatePublished - Nov 2011
Externally publishedYes

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Chlamydia trachomatis
Plasmids
Monoclonal Antibodies
Proteins
Chlamydia muridarum
Chlamydophila psittaci
Chlamydophila pneumoniae
Antibody Specificity
Antibodies
Hybridomas
Affinity Chromatography
Fluorescent Antibody Technique
Western Blotting

Keywords

  • Chlamydia trachomatis
  • Immunological characteristics
  • Monoclonal antibody
  • PORF5 plasmid protein

ASJC Scopus subject areas

  • Immunology
  • Microbiology
  • Virology

Cite this

Purification and immunological characteristics of monoclonal antibody 2H4 against Chlamydia trachomatis pORF5 plasmid protein. / Li, Zhong Yu; Wu, Yi Mou; Huang, Qiu Lin; Su, Sheng Mei; Zhou, Zhou; Chen, Chao Qun; Zhou, Hui; Zhong, Guangming.

In: Chinese Journal of Microbiology and Immunology (China), Vol. 31, No. 11, 11.2011, p. 1041-1045.

Research output: Contribution to journalArticle

Li, Zhong Yu ; Wu, Yi Mou ; Huang, Qiu Lin ; Su, Sheng Mei ; Zhou, Zhou ; Chen, Chao Qun ; Zhou, Hui ; Zhong, Guangming. / Purification and immunological characteristics of monoclonal antibody 2H4 against Chlamydia trachomatis pORF5 plasmid protein. In: Chinese Journal of Microbiology and Immunology (China). 2011 ; Vol. 31, No. 11. pp. 1041-1045.
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abstract = "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.",
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AU - Wu, Yi Mou

AU - Huang, Qiu Lin

AU - Su, Sheng Mei

AU - Zhou, Zhou

AU - Chen, Chao Qun

AU - Zhou, Hui

AU - Zhong, Guangming

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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

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