[Screening and identification of human anti-c-Met Fab from a phage antibody library].
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.
Sun
Hui
H
Department of Pathology, Nanjing Medical University, Nanjing 210029, China.
Wan
Jia-Yi
JY
Zhu
Xiao-Juan
XJ
Jiao
Yong-Jun
YJ
Zhu
Jin
J
Feng
Zhen-Qing
ZQ
Liu
Zheng
Z
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
D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D019859:Proto-Oncogene Proteins c-met