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| Preparation and specificity analysis of monoclonal antibody against 6 kD early secretory antigenic target of Mycobacterium tuberculosis |
| NING Huanhuan1 REN Rui1 KANG Jian1 BAI Lu2 LU Yanzhi1 XIE Yanling2 ZHANG Fanglin1 BAI Yinlan1 |
| 1.Department of Microbiology and Pathogen Biology, School of Preclinical Medicine, Air Force Medical University, Shaanxi Province, Xi’an 710032, China; 2.School of Life Sciences, Yan’an University, Shaanxi Province, Yan’an 716000, China |
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Abstract Objective To prepare a monoclonal antibody (mAb) against 6 kD early secretory antigenic target (ESAT-6) of Mycobacterium tuberculosis and to analyze its specificity. Methods ESAT-6 was purified by affinity chromatography and immunized mice with the protein. Splenocytes from ESAT-6 immunized mice were fused with myeloma cells, and hybridoma cell lines secreting mAb were screened. mAb was purified from ascites. The subclass and relative affinity of mAb were assayed by enzyme-linked immunosorbent assay. The specificity of the mAb was analyzed by Western blot. Results The hybridoma cell line secreting ESAT-6 antibody was obtained and mAb was purified. The subclass of ESAT-6 mAb was IgG1 with a relative affinity of 3.9×10-5. The mAb specifically recognizes ESAT-6 in Mtb. Conclusion The preparation of mAb against ESAT-6 lays a foundation for the application of ESAT-6 in the diagnosis and treatment of tuberculosis and its mechanism.
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