Production and characterization of monoclonal antibodies to Mycobacterium tuberculosis.

Senthil Kumar, K.S.; Alamelu Raja; Uma Devi, K.R.; Paranjape, R.S.

Indian Journal of Medical Research; 2000; 112; 37-46.          

Background and objectives : Monoclonal antibdoies (MAbs) against Mycobacterium tuberculosis H37Rv culture filtrate (CF) were raised by immunizing BALB/c mice and characterization was done. Attempts have been directed towards identifying mycobacterial antigens in biological fluids by employing polyclonal and monoclonal antibodies specific for M. tuberculosis . Immunohistologic studies, using MAbs for the localization of whole or fragmented bacilli in the biopsy specimens were also carried out.

Methods : Intrasplenic IS and intraperitoneal IP routes of immunization, were compared. The MAbs were characterized for their isotype, binding specificity, nature of binding epitope, reactivity in immunoassays etc.

Results : IS and IP routes of immunization, were compared and IP was found superior. Ten MAbs designated TRC 1-10 were produced. Of these, 7 MAbs, TRC 1-7 reacted with the 30-31 kDa doublet (antigen 85 complex), TRC 8 with 12 kDa in addition to 30/31 kDa and TRC 9 and 10 with the 24 and 12 kDa antigens respectively. Six MAbs were classified as broadly cross reactive and 2 showed limited cross reactivity. TRC 8 and 10 showed species specificity. Employing TRC 8 in sandwich ELISA, antigen was detected in sera from 17 of 25 pulmonary tuberculosis patients and 3 of 20 controls. TRC 8 was found to be useful in detecting antigen specifically in M. tuberculosis and M. leprae infected tissues, by immunoperoxidase staining.

Interpretation & conclusion : TRC 8 was found to be restricted in its reactivity to M. tuberculosis complex and M. leprae . TRC 8 may prove useful in immuno-diagnosis of tuberculosis.

Key words : Enzyme linked immunosorbent assay (ELISA) - immuno-peroxidase staining - intrasplenic immunization (IS) - Mycobacterium tuberculosis - monoclonal antibodies (MAbs) - sandwich ELISA

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