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

Area of Science:

• Oncology
• Molecular Biology
• Immunotherapy

Background:

• The receptor tyrosine kinase (RTK) Met, activated by hepatocyte growth factor (HGF), plays a critical role in cell growth, migration, and survival.
• Dysregulation of c-Met signaling is implicated in colorectal cancer (CRC) progression, driving proliferation, invasion, and angiogenesis.
• Targeting the extracellular domain of c-Met offers a potential therapeutic strategy to block aberrant signaling in cancer.

Purpose of the Study:

• To develop novel molecular inhibitors targeting the extracellular domain of the c-Met receptor.
• To isolate and characterize single chain variable fragments (scFv) with specific binding affinity for c-Met.
• To evaluate the therapeutic potential of selected c-Met specific scFvs in inhibiting colorectal cancer cell growth and inducing apoptosis.

Main Methods:

• Screening of Tomlinson I+J scFv phage display libraries against a synthetic oligopeptide from the c-Met extracellular domain.
• Selection and characterization of c-Met specific scFvs using immune techniques.
• Assessment of scFv binding affinity to c-Met and their effect on proliferation and apoptosis of human colorectal carcinoma HCT-116 cells.

Main Results:

• Three specific c-Met targeting scFvs (ES1, ES2, ES3) were successfully isolated after five rounds of panning.
• The selected scFvs demonstrated specific binding to the c-Met receptor.
• ES1, ES2, and ES3 significantly inhibited proliferation and induced apoptosis in HCT-116 cells, with cell death rates of 46.0%, 25.5%, and 37.8%, respectively.

Conclusions:

• Specific c-Met targeting scFvs have been successfully developed and characterized.
• These scFvs exhibit potent anti-proliferative and pro-apoptotic effects on colorectal cancer cells.
• The isolated c-Met scFvs hold significant therapeutic potential for the development of novel immunotherapies against colorectal cancers.