title: [Cloning and expressing of tissue inhibitor of metalloproteinases I gene fragment and preparation of monoclonal antibodies against the recombinant protein].
abstract: OBJECTIVE
  To prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein.
  
  
  METHODS
  TIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot were used to detect specificity of mAbs.
  
  
  RESULTS
  The prokaryotic plasmid expressing the recombinant protein was constructed, and the TIMP-I recombinant protein was expressed and purified. Four hybridoma cell lines that secreted anti-TIMP-I mAbs were obtained. 3 of 4 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with TIMP-I protein.
  
  
  CONCLUSIONS
  The TIMP-I recombinant protein is highly purified and has strong antigenicity. The anti- TIMP-I mAbs were prepared successfully.
authors: Medical University of PLA, Beijing 100853, China.
authors: Jun
authors: J
authors: Hou
authors: Jian
authors: J
authors: Zhang
authors: Jing-Xia
authors: JX
authors: Guo
authors: Lin
authors: L
authors: Cheng
authors: Jing
authors: J
authors: Zhao
authors: Jia
authors: J
authors: Liu
authors: Jun
authors: J
authors: Xu
authors: Ai-Xia
authors: AX
authors: Liu
authors: Yong-Ji
authors: YJ
authors: Song
authors: Pan-Yong
authors: PY
authors: Mao
authors: Bo-An
authors: BA
authors: Li
authors: Yuan-Li
authors: YL
authors: Mao
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D003001:Cloning, Molecular; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1