title: [Screening and identification of human anti-c-Met Fab from a phage antibody library].
abstract: OBJECTIVE
  To screen anti-c-Met Fab from a phage antibody library and identify its binding activity.
  
  
  METHODS
  The expression of c-Met of HCC lines was identified by Western blot and immunofluorescence. Antibodies against c-Met were screened with immobilized antigen. After five rounds of panning, 30 randomly selected clones were identified by phage ELISA to select specific clones with high affinity. The positive clones were selected for Fab soluble expression in TOP10F and the binding activities were analysed in HCC lines.
  
  
  RESULTS
  c-Met expressed in HCC membrane was confirmed by Western blot and immunofluorescence. A Fab fragment named AM2-26 with fine activity to c-Met was selected. AM2-26 binding specificity was confirmed by IP, FACS and immunofluorescence.
  
  
  CONCLUSIONS
  The anti-c-Met Fab binding to c-Met in HCC provides a promising candidate for the biotherapy of hepatoma.
authors.affiliation: Department of Pathology, Nanjing Medical University, Nanjing 210029, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Sun
authors.forename: Jia-Yi
authors.initials: JY
authors.lastname: Wan
authors.forename: Xiao-Juan
authors.initials: XJ
authors.lastname: Zhu
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Jiao
authors.forename: Jin
authors.initials: J
authors.lastname: Zhu
authors.forename: Zhen-Qing
authors.initials: ZQ
authors.lastname: Feng
authors.forename: Zheng
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D015723:Gene Library; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000276:immunology; D019151:Peptide Library; D019859:Proto-Oncogene Proteins c-met / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology
chemical_list: D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D019859:Proto-Oncogene Proteins c-met