Experimental Research Of Screening Biomarkers In The Sera Of PLC With Phage Random Peptide Library

Background and Objective: Until now, Primary Liver Cancer,(PLC) is still difficult to obtain an early diagnose. However, new biomarkers can be recognized in the sera of the patients.Detection of these biomarkers may provide a means for sensitive and specific diagnosis.AFP is one of the most important biomarkers in the PLC,But only 70% of PLC patients have a high serum concentration of AFP, It can have a high concentration in some benign diseases too. New biomarkers with better specific are needed.Screening phage display libraries of peptides is a powerful technology for selecting polypeptides or proteins.Fragments of foreign peptides or proteins are expressed as fusion proteins displayed on the phage surface,keeping their relatively independent spatial structure and biological acti- vity.Through a immunoscreening of phage random peptide library with target protein, we obtain the single phage clone and the specific peptide. Then the peptide can be sequenced to obtain the corresponding information,such as the structure and the function.So we use the technology to search new antigen peptides and provide new markers for the diagnosis and prognosis of PLC.Methods: 1.Through the pre-adsorption with the normal serum, non-combination phage were obtained and adsorbed with the serum of PLC. The positive clones were obtained through three rounds of biopanning.2.The reactivity of each clone bound to the serum was examined by ELISA. Finally, we selected the clones could bind to the sera of PLC patients.3.The positive clones were sequenced with -96â…¢primers, we obtain the protein and the peptides.Results: The eluted phages were enriched nearly to 100 fold through three rounds of biopanning. From the clones selected randomly from the third round biopanning, we selected the best 3 clones could bind to the sera from PLC patients. The peptide may be new biomarkers of PLC.Conclusion: The peptides from immunoscreening of phage random peptide library may supply candidate biomarkers of diagnose, combined detection and immunotherapy of PLC.