Screening and identification of human anti-c-Met Fab from a phage antibody library

To screen anti-c-Met Fab from a phage antibody library and identify its binding activity. 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 cl...

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Published inZhonghua gan zang bing za zhi Vol. 16; no. 7; p. 505
Main Authors Sun, Hui, Wan, Jia-Yi, Zhu, Xiao-Juan, Jiao, Yong-Jun, Zhu, Jin, Feng, Zhen-Qing, Liu, Zheng
Format Journal Article
LanguageChinese
Published China 01.07.2008
Subjects
Antibodies - immunology
Antibodies - 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
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Summary:To screen anti-c-Met Fab from a phage antibody library and identify its binding activity. 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. 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. The anti-c-Met Fab binding to c-Met in HCC provides a promising candidate for the biotherapy of hepatoma.
ISSN:1007-3418