| Production and Study on the Specificity of Monoclonal Antibodies to Pressate Extract Antigen of Mycobacterium tuberculosis |
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Yoon Hoh Kook1, Myung Je Cho1, Eung Soo Hwang1, Ik Sang Kim1, Chang Yong Cha1, Woo Hyun Chang1, Seung Hoon Lee1, Sang Jae Kim2, Yong Chol Han3 |
1Dept. of Microbiology, College of Medicine, Seoul National University, Seoul, Korea
2Korean Institutes of Tuberculosis , Korean National Tuberculosis Association, Seoul, Korea
3Dept. of Internal Medicine, College of Medicine, Seoul National University, Seoul, Korea |
| Mycobacterium tuberculosis 파쇄 추출항원에 대한 단세포군 항체의 생산 및 그 특성에 관한 연구 |
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국윤호1, 조명제1, 황응수1, 김익상1, 차창룡1, 장우현1, 이승훈1, 김상재2, 한용철3 |
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| Abstract |
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Pressate extract of M. tuberculosis was used as antigen for immunization of Balb/c mouse to obtain spleen cells producing antibodies to M. tuberculosis. Antibody producing spleen cells were prepared 3 days after the last . challenge and then fused with HGPRT deficient, nonsecreting mouse myeloma cell line, P3-X 63-Ag 8.653. Primary screening by enzyme linked immunosorbent assay(ELISA) was performed 2 weeks after fusion to determine antibody secreting hybridom cells in 96-we11 culture plates, and among 576 wells, 124 wells (21. 5%) were positive. These hybridom cells were cloned by limiting dilution method and 5 celllines (TB-1 , TB-2, TB-3, TB-4 and TB-5) , were expanded to obtain culture supernatants containing antibodies reacting to M. tuberculosis antigen. These 5 moüoclonal antibodies were tested for their specificity toM. tuberculosis extract antigen by antibody absorption tests. All showed below than 50% of absorption when mixed with each extract antigen from M. bovis, M. aviltm, and M. fortuitum and no reactivities on ELl SA plates coated with these three antigens. Antibodies produced by TB-2 and TB-3 cell lines were determined to be more specific to TE antigen than those from TB-l , TB-4, and TB-5 hybridom, cells, These hybridoma cells were injected into the peritoneal cavity of Balb/c mice piletreated with pristane to induce tumor. 7 days after injection of the hybridoma cells, the antibody titer of ascitic fluid were determined by ELISA. All showed 2 or 3 times higher optical density in ELISA than those of hybridoma cell culture fluid. |
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