Keywords

Deoxynivalenol (Don), Phage Display Random Peptide Library, Enzyme-Linked Immunoassay, Wheat Scab

Abstract

In order to obtain a specific peptide with high affinity for the toxin deoxynivalenol (DON), the DON artificial antigen DON-HG-BSA was prepared using the butylboronic acid blocking method and used as a target molecule to screen the phage peptide library. Positive clones were subjected to DNA sequence determination and amino acid derivation, and affinity specificity verification was performed. The results showed that after 4 rounds of affinity peptide screening and enzyme-linked immunoassay identification, 15 positives with an OD ratio of DON artificial antigen-coated wells and bovine serum albumin-coated wells greater than 3.8 were obtained from the M13 phage random 7-peptide library. Among them, 10 positive clones that bind with high affinity to DON artificial antigen were sequenced and a total of 4 polypeptide sequences were deduced. Verified by the antigen gradient enzyme-linked immunoassay method, the correlation between the concentration of the peptide MAPGWVP and DON artificial antigen is as high as 0.99, and it has high affinity for the artificial antigen. The acquisition of this peptide will lay the foundation for further development of efficient DON diagnostic kits.

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Original research done by Xing Jincheng, Zhang Xu, Ma Hongxiang, Xu Hai, Hou Jibo

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