Construction Of Epitope Nonapeptide Library And Screening Of Specific Binding Peptides For Human Type Iv Collagenase
Keywords
Epitope Peptide Library, Nonapeptide, Type Iv Collagenase, Affinity Screening
Abstract
The synthetic random sequence DNA fragment encoding nonapeptide was cloned into the filamentous phage expression vector FUSE5. After multiple electroporations, transformation and expression, the random sequence nonapeptide epitope peptide was obtained by fusion of the peptide fragment with the phage pIII protein and displayed on the surface of the phage. library. Library capacity reaches 10 10 clones. Using type IV collagenase as the target protein, affinity purification screening mode was used to screen out type IV collagenase-binding peptides. Further ELISA testing screened out 20 positive clones that specifically bind to type IV collagenase. Sequence analysis found that one group of peptides contained the common sequence WDXXD, and one group contained the common sequence WVGXXR. The sequence of WDXXD is homologous to the variable region sequence of type IV collagenase single-chain antibody. The results show that the peptide library is a powerful tool for screening protein-specific binding peptides. The construction of the epitope nonapeptide library and the establishment of the screening method laid the foundation for further applications in screening specific binding peptides with high affinity.
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Original research was done by Li Chuanzhao, Yang Wending, Liu Yule, Wang Dawei, Zhao Aimin, Tian Bo
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