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Updated: Jun 18, 2026
Separation and Fractionation of Culture Filtrate Proteins (CFPs) from Mycobacterium tuberculosis
Published on: July 11, 2025
[Preparation and characterization of monoclonal antibodies specific for CFP-10 protein of Mycobacterium
Jun-Song Shen1, Hui Zhang, Cheng-Quan Zhang
1Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China.
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.
