Biopanning of Polypeptides Binding to Bovine Ephemeral Fever Virus G Protein from Phage Display Peptide Library

Overview

Journal BMC Vet Res

Publisher Biomed Central

Specialty Veterinary Medicine

Date 2018 Jan 6

PMID 29301517

Citations 22

Authors

Peili Hou

Guimin Zhao

Chengqiang He

Hongmei Wang

Hongbin He

Affiliations

Soon will be listed here.

Abstract

Background: The bovine ephemeral fever virus (BEFV) glycoprotein neutralization site 1 (also referred as G protein), is a critical protein responsible for virus infectivity and eliciting immune-protection, however, binding peptides of BEFV G protein are still unclear. Thus, the aim of the present study was to screen specific polypeptides, which bind BEFV G protein with high-affinity and inhibit BEFV replication.

Methods: The purified BEFV G was coated and then reacted with the M13-based Ph.D.-7 phage random display library. The peptides for target binding were automated sequenced after four rounds of enrichment biopanning. The amino acid sequences of polypeptide displayed on positive clones were deduced and the affinity of positive polypeptides with BEFV G was assayed by ELISA. Then the roles of specific G-binding peptides in the context of BEFV infection were analyzed.

Results: The results showed that 27 specific peptide ligands displaying 11 different amino acid sequences were obtained, and the T18 and T25 clone had a higher affinity to G protein than the other clones. Then their antiviral roles of two phage clones (T25 and T18) showed that both phage polypeptide T25 and T18 exerted inhibition on BEFV replication compared to control group. Moreover, synthetic peptide based on T18 (HSIRYDF) and T25 (YSLRSDY) alone or combined use on BEFV replication showed that the synthetic peptides could effectively inhibit the formation of cytopathic plaque and significantly inhibit BEFV RNA replication in a dose-dependent manner.

Conclusion: Two antiviral peptide ligands binding to bovine ephemeral fever virus G protein from phage display peptide library were identified, which may provide a potential research tool for diagnostic reagents and novel antiviral agents.

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