Preparation and characterization of monoclonal antibodies specific for CFP-10 protein of Mycobacterium tuberculosis.
To prepare monoclonal antibodies (mAb) against CFP-10 protein of Mycobacterium tuberculosis. BALB/c mice were immunized with the purified His-CFP-10 expressed in BL21 (DE3)-pET-30a(+)-lhp. With the purified GST-CFP-10 as detecting antigen, mAb-produced hybridoma cells against CFP-10 were screened by...
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Published in | Xi bao yu fen zi mian yi xue za zhi Vol. 25; no. 12; p. 1143 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | Chinese |
Published |
China
01.12.2009
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Subjects | |
Online Access | Get more information |
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Summary: | To prepare monoclonal antibodies (mAb) against CFP-10 protein of Mycobacterium tuberculosis.
BALB/c mice were immunized with the purified His-CFP-10 expressed in BL21 (DE3)-pET-30a(+)-lhp. With the purified GST-CFP-10 as detecting antigen, mAb-produced hybridoma cells against CFP-10 were screened by indirect ELISA. The specificity of the mAbs were characterized by indirect ELISA, Dot-ELISA and Western blot.
Two hyridoma cell lines secreting mAbs against CFP-10 named 6E8, 2E7 were obtained. The immunoglobulin subclasses of 2 mAbs were IgG1 and IgG2b respectively, and the ELISA titers of 2 mAbs ascitic fluids were 1:1 000 000, 1:1 024 000 respectively. In Dot-ELISA test, the 2 mAbs could only react with BL21 (DE3)-pET-30a(+)-lhp, BL21-pGEX-6P-1-lhp, which expressed His-CFP-10, GST-CFP-10, respectively. Western blot analysis confirmed that the 2 mAbs could only react with CFP-10 protein.
Two mAbs specific to CFP-10 protein of Mycobacterium tuberculosis were obtained, which may have important application value in |
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ISSN: | 1007-8738 |