| Production and Characterization of Monoclonal Antibodies Reactive to PPD of Mycobacterium Tuberculosis |
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Y. S. Shim1, Y. C. Han1, M. J. Cho2, C. Y. Cha2 |
1Department of Internal Medicine, College of Medicine, Seoul National University, Seoul, Korea
2Department of microbiology, College of Medicine, Seoul National University, Seoul, Korea |
| 결핵균 PPD(RT40)에 반응하는 단세포군 항체의 생산 및 그 특성에 관한 연구 |
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심영수1, 한용철1, 조명제2, 차창룡2 |
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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 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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