Screening and identification of human anti-c-Met Fab from a phage antibody library.
- Author:
Hui SUN
1
;
Jia-Yi WAN
;
Xiao-Juan ZHU
;
Yong-Jun JIAO
;
Jin ZHU
;
Zhen-Qing FENG
;
Zheng LIU
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies; immunology; isolation & purification; Cell Line, Tumor; Cloning, Molecular; Gene Library; Humans; Immunoglobulin Fab Fragments; immunology; Immunoglobulin Variable Region; immunology; Peptide Library; Proto-Oncogene Proteins c-met; immunology; Recombinant Fusion Proteins; immunology
- From: Chinese Journal of Hepatology 2008;16(7):505-508
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screen anti-c-Met Fab from a phage antibody library and identify its binding activity.
METHODSThe 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.
RESULTSc-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.
CONCLUSIONThe anti-c-Met Fab binding to c-Met in HCC provides a promising candidate for the biotherapy of hepatoma.
