Keywords

Endotoxins, Phage Display Random Peptide Library, Peptides, Bioinformatics

Abstract

Objective To screen peptide sequences that bind to endotoxin from a phage display random peptide library and identify them. Methods Four rounds of affinity screening were conducted on a phage display random twelve-peptide library using endotoxin lipopolysaccharide (LPS) as the target molecule, and phage clones that combined with LPS were obtained, and further confirmed by binding experiments and clone plaque inhibition experiments. Select clones with strong binding ability for DNA sequencing, deduce the presented polypeptide sequence, and use bioinformatics software for polypeptide sequence analysis and homology analysis. Results After 4 rounds of affinity screening, 86 clones were screened from the phage display random twelve peptide library, and 12 clones with strong binding ability were selected for DNA sequence sequencing and bioinformatics analysis. Combined with the phage display random seven clones of this project team, The peptide library screening results deduced that the presented polypeptide sequence is HWQWPHWSPPP (named P11 peptide). Searching relevant databases found that this peptide sequence has not been patented, and about 908 proteins in the body match its structure, including sites that interact with LPS. Conclusion By panning the phage display random peptide library, high-affinity polypeptides that bind to LPS were obtained, which provided experimental basis and structural basis for further directed evolution research using these polypeptides as precursors.

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Original research done by Yang Haijie, Duan Guangjie, Liu Yousheng, Chen Rui, Yu Dewen, Yang Yu

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