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

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2009 Dec;25(12):1143-5.
[Article in Chinese]

Abstract

Aim: To prepare monoclonal antibodies (mAb) against CFP-10 protein of Mycobacterium tuberculosis.

Methods: 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.

Results: 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.

Conclusion: Two mAbs specific to CFP-10 protein of Mycobacterium tuberculosis were obtained, which may have important application value in further studies on diagnosis and pathogenesis.

MeSH terms

  • Animals
  • Antibodies, Monoclonal* / immunology
  • Blotting, Western
  • Enzyme-Linked Immunosorbent Assay
  • Hybridomas / immunology
  • Mice, Inbred BALB C
  • Mycobacterium tuberculosis* / immunology

Substances

  • Antibodies, Monoclonal