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KMID : 0383819870340010013
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Tuberculosis and Respiratory Diseases
1987 Volume.34 No. 1 p.13 ~ p.26
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Production and Characterization of Monoclonal Antibodies Reactive to PPD of Mycobacterium Tuberculosis
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Abstract
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Using hybridoma technique, the analysis of antigenicity of tuberculin purified protein
derivative(PPD) antigen and the identification of the species-specific antigen of
Mycobacterirm tuberculosis were attempted. Moncolonal antibodies to tuberculin PPD
were produced by the fusion of BALB/c mouse spleen cells with BALB/c mouse
myeloma (P3×63-Ag8. v653) cells.
Fifteen monoclonal antibodies were characterized as to their reactivity with tuberculin
PPD antigen and Mycobacterirm 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 mycobacterial 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 Mycobacterirm tuberculosis antigen when
tested against the sonicate and fractionate antigens of 5 mycobacterial species.
Nine antigenic determinants of Mycobacterirm 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 Mycobacterirm tuberculosis. 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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