Preparation and characterization of monoclonal antibodies specific for CFP-10 protein of Mycobacterium tuberculosis.

To prepare monoclonal antibodies (mAb) against CFP-10 protein of Mycobacterium tuberculosis. BALB/c mice were immunized with the purified His-CFP-10 expressed in BL21 (DE3)-pET-30a(+)-lhp. With the purified GST-CFP-10 as detecting antigen, mAb-produced hybridoma cells against CFP-10 were screened by...

Full description

Saved in:
Bibliographic Details
Published inXi bao yu fen zi mian yi xue za zhi Vol. 25; no. 12; p. 1143
Main Authors Shen, Jun-Song, Zhang, Hui, Zhang, Cheng-Quan, Pan, Zhi-Ming, Yin, Yue-Lan, Jiao, Xin-An
Format Journal Article
LanguageChinese
Published China 01.12.2009
Subjects
Animals
Antibodies, Monoclonal - immunology
Blotting, Western
Enzyme-Linked Immunosorbent Assay
Hybridomas - immunology
Mice, Inbred BALB C
Mycobacterium tuberculosis - immunology
Online AccessGet more information

Cover

More Information
Summary:To prepare monoclonal antibodies (mAb) against CFP-10 protein of Mycobacterium tuberculosis. BALB/c mice were immunized with the purified His-CFP-10 expressed in BL21 (DE3)-pET-30a(+)-lhp. With the purified GST-CFP-10 as detecting antigen, mAb-produced hybridoma cells against CFP-10 were screened by indirect ELISA. The specificity of the mAbs were characterized by indirect ELISA, Dot-ELISA and Western blot. Two hyridoma cell lines secreting mAbs against CFP-10 named 6E8, 2E7 were obtained. The immunoglobulin subclasses of 2 mAbs were IgG1 and IgG2b respectively, and the ELISA titers of 2 mAbs ascitic fluids were 1:1 000 000, 1:1 024 000 respectively. In Dot-ELISA test, the 2 mAbs could only react with BL21 (DE3)-pET-30a(+)-lhp, BL21-pGEX-6P-1-lhp, which expressed His-CFP-10, GST-CFP-10, respectively. Western blot analysis confirmed that the 2 mAbs could only react with CFP-10 protein. Two mAbs specific to CFP-10 protein of Mycobacterium tuberculosis were obtained, which may have important application value in
ISSN:1007-8738