Tuberc Respir Dis.  1987 Mar;34(1):13-26. 10.4046/trd.1987.34.1.13.

Production and Characterization of Monoclonal Antibodies Reactive to PPD of Mycobacterium Tuberculosis

Abstract

Using hybridoma technique, the analysis of antigenicity of tuberculin purified protein derivative (PPD) antigen and the identification of the species-specific antigen of Mycobacterium tuberculosis were attempted. Moncolonal antibodies to tuberculin PPD were produced by the fusion of BALB/c mouse spleen cells with BALB/c mouse myeloma (P3X63-Ag8. v653) cells. Fifteen monoclonal antibodies were characterized as to their reactivity with tuberculin PPD antigen and Mycobacterium tuberculosis sonicate (TE) antigen by enzyme-linked immunosorbent assay. Among these, 8 monoclonal antibodies reacted with both tuberculin PPD antigen and TE antigen. All the antibodies were of IgG class. The subclass of antibody MPT-1 was IgG 2a and others were IgG 1. The reactivity of the antibodies with 8 kinds of mycobacteria PPD were examined by enzyme-linked immunosorbent assay. Antibody MP-2, MP-5 & MPT-1 reacted selectively with tuberculin PPD antigen- Antibody MPT-6 cross-reacted weakly with 5 atypical mycobacterial PPD antigens. Among 8 antibodies, reacting with both tuberculin PPD antigen and TE antigen, MPT-1 and MPT-6 were found to be specific of Mycobacterium tuberculosis antigen when tested against the sonicate and fractionate antigens of 5 mycobacterial species. Nine antigenic determinants of Mycobacterium tuberculosis were identified by the reaction pattern of the antibody with PPD, sonicate and fractionate antigens. PPD-Antigen 1 was present only in tuberculin PPD and PPD-Antigen 4 was the specific antigen of Myco. bacte서μm t:μberα , lo.sis. It is suggested that MPT-1 is the specific monoclonal antibody reacting with species-specific antigenic determinant. Also , this monoclonal antibody would be a valuable diagnostic reagent as well as a tool for the purification of specific antigen.

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