[Preparation and characterization of monoclonal antibodies specific for CFP-10 protein of Mycobacterium tuberculosis.].
OBJECTIVE
To prepare monoclonal antibodies (mAb) against CFP-10 protein of Mycobacterium tuberculosis.


METHODS
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.


RESULTS
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.


CONCLUSIONS
Two mAbs specific to CFP-10 protein of Mycobacterium tuberculosis were obtained, which may have important application value in further studies on diagnosis and pathogenesis.
Shen
Jun-Song
JS
Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China.
Zhang
Hui
H
Zhang
Cheng-Quan
CQ
Pan
Zhi-Ming
ZM
Yin
Yue-Lan
YL
Jiao
Xin-An
XA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000276:immunology
D000911:Antibodies, Monoclonal