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Novel p16 binding peptide development for p16-overexpressing cancer cell detection using phage display
Numfon Khemthongcharoen1, Athisake Ruangpracha, Pongsak Sarapukdee
1NECTEC, National Science and Technology Development Agency (NSTDA), Pathumthani, 12120, Thailand; Advanced Imaging Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
Researchers identified novel peptides that bind to p16INK4a (p16), a key biomarker for human papillomavirus (HPV)-related cancers. These p16 binding peptides show potential for detecting cancer cells overexpressing p16.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Biomarkers
Background:
- Protein p16INK4a (p16) is a crucial biomarker for diagnosing human papillomavirus (HPV)-related cancers.
- Developing specific tools for p16 detection is vital for accurate cancer diagnosis and management.
Purpose of the Study:
- To identify novel peptides that specifically bind to the p16 protein using phage display technology.
- To evaluate the binding affinity, specificity, and selectivity of the identified peptides for p16.
Main Methods:
- Phage display selection was employed using a random heptamer library screened against purified recombinant p16 protein.
- Binding affinity was assessed using ELISA and fluorescence imaging.
- Peptide specificity and selectivity were evaluated through peptide blocking experiments and fluorescence imaging of cell lines.
Main Results:
- Five candidate p16-binding peptides were isolated.
- Three peptides demonstrated significant binding affinity and selectivity for p16-overexpressing cancer cells (CaSki) compared to normal cells (HUF).
- The peptide 'Ser-His-Ser-Leu-Leu-Ser-Ser' exhibited the best binding score, did not interfere with p16 function, and blocked monoclonal antibody interaction.
Conclusions:
- Novel p16-binding peptides were successfully identified using phage display.
- The peptide 'Ser-His-Ser-Leu-Leu-Ser-Ser' shows high potential for detecting p16-overexpressing cells and can be a valuable tool in cancer diagnostics.
