title: [Preparation and characterization of monoclonal antibodies specific for CFP-10 protein of Mycobacterium tuberculosis.].
abstract: 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.
authors: Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou 225009, China.
authors: Jun-Song
authors: JS
authors: Shen
authors: Hui
authors: H
authors: Zhang
authors: Cheng-Quan
authors: CQ
authors: Zhang
authors: Zhi-Ming
authors: ZM
authors: Pan
authors: Yue-Lan
authors: YL
authors: Yin
authors: Xin-An
authors: XA
authors: Jiao
mesh_terms: 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
chemical_list: D000911:Antibodies, Monoclonal