Isolation and characterization of anti c-met single chain fragment variable (scFv) antibodies

• Published in: Journal of immunotoxicology 2017-01, Vol.14 (1), p.23-30
• Main Authors: Qamsari, Elmira Safaie, Sharifzadeh, Zahra, Bagheri, Salman, Riazi-Rad, Farhad, Younesi, Vahid, Abolhassani, Mohsen, Ghaderi, Sepideh Safaei, Baradaran, Behzad, Somi, Mohammad Hossein, Yousefi, Mehdi
• Format: Article
• Language: English
• Subjects: Angiogenesis; Annexin V; Antibodies; Antibodies, Monoclonal - isolation & purification; Antibodies, Monoclonal - pharmacology; Antigens; Apoptosis; Apoptosis - drug effects; c-Met; c-Met protein; Carcinogenesis; Cell death; Cell Growth Processes - drug effects; Cell surface; Cell Surface Display Techniques; Colorectal cancer; Colorectal carcinoma; Colorectal Neoplasms - immunology; Colorectal Neoplasms - metabolism; Colorectal Neoplasms - therapy; E coli; Embryogenesis; Growth factors; HCT116 Cells; Hepatocyte growth factor; Hepatocyte Growth Factor - metabolism; HGF; Humans; Immunoglobulins; Immunology; Immunotherapy - methods; Invasiveness; Kinases; Laboratories; Medical prognosis; Metastasis; Panning; Phage display; Prostate; Protein-tyrosine kinase receptors; Proto-Oncogene Proteins c-met - genetics; Proto-Oncogene Proteins c-met - immunology; Proto-Oncogene Proteins c-met - metabolism; Research centers; scFv; Single-Chain Antibodies - isolation & purification; Single-Chain Antibodies - pharmacology; Tumors
• ISSN: 1547-691X; 1547-6901
• DOI: 10.1080/1547691X.2016.1251512

The receptor tyrosine kinase (RTK) Met is the cell surface receptor for hepatocyte growth factor (HGF) involved in invasive growth programs during embryogenesis and tumorgenesis. There is compelling evidence suggesting important roles for c-Met in colorectal cancer proliferation, migration, invasion, angiogenesis, and survival. Hence, a molecular inhibitor of an extracellular domain of c-Met receptor that blocks c-Met-cell surface interactions could be of great thera-peutic importance. In an attempt to develop molecular inhibitors of c-Met, single chain variable fragment (scFv) phage display libraries Tomlinson I + J against a specific synthetic oligopeptide from the extracellular domain of c-Met receptor were screened; selected scFv were then characterized using various immune techniques. Three c-Met specific scFv (ES1, ES2, and ES3) were selected following five rounds of panning procedures. The scFv showed specific binding to c-Met receptor, and significantly inhibited proliferation responses of a human colorectal carcinoma cell line (HCT-116). Moreover, anti- apoptotic effects of selected scFv antibodies on the HCT-116 cell line were also evaluated using Annexin V/PI assays. The results demonstrated rates of apoptotic cell death of 46.0, 25.5, and 37.8% among these cells were induced by use of ES1, ES2, and ES3, respectively. The results demonstrated ability to successfully isolate/char-acterize specific c-Met scFv that could ultimately have a great therapeutic potential in immuno-therapies against (colorectal) cancers.