Design and preliminary application of affinity peptide based on the structure of the porcine circovirus type II Capsid (PCV2 Cap)

Junfang Hao; Fangyu Wang; Guangxu Xing; Yunchao Liu; Ruiguang Deng; Hao Zhang; Anchun Cheng; Gaiping Zhang

doi:10.7717/peerj.8132

Design and preliminary application of affinity peptide based on the structure of the porcine circovirus type II Capsid (PCV2 Cap)

PeerJ. 2019 Dec 5:7:e8132. doi: 10.7717/peerj.8132. eCollection 2019.

Authors

Junfang Hao^#^{1

2}, Fangyu Wang^#², Guangxu Xing², Yunchao Liu², Ruiguang Deng², Hao Zhang^{2

3}, Anchun Cheng¹, Gaiping Zhang^{1

2

3}

Affiliations

¹ Research Center of Avian Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China.
² Henan Key Laboratory for Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou, China.
³ College of Life Science, Henan Agricultural University, Zhengzhou, China.

^# Contributed equally.

Abstract

Background: Affinity peptides, as a core part of affinity chromatography, play an important role in the purification of target molecules.

Methods: Here we describe the use of molecular docking technology for virtual screening of affinity peptides that specifically recognize the PCV2 Cap protein for the first time. Thirteen candidate peptides with high scores were obtained and then further characterized. Experimentally, the affinity and sensitivity of the peptides studied were identified by ELISA and LSPR, respectively. In order to investigate the purification effect of a selected peptide (L11) for the recombinant PCV2 Cap protein, it was coupled to NHS agarose magnetic beads as an affinity adsorbent (NaMB-L11); and the ligand density of the affinity adsorbent and pH value in the purification of the recombinant PCV2 Cap protein were optimized.

Results: Our data showed that the peptide L11- DYWWQSWE has the smallest K_D = 103 nM with higher specificity for PCV2 Cap protein recognition. The NaMB-L11 affinity adsorbent yielded a purified Cap sample with 98% purity at 90% recovery in a single step.

Conclusion: Based on the structure, we obtained a high affinity peptide L11 binding to the PCV2 Cap protein by molecular docking technology. It not only provides a theoretical basis for the design of PCV2 Cap affinity peptide, but a new method for the purification of the PCV2 Cap protein.

Keywords: Affinity peptides; Molecular docking virtual screening; PCV2 Cap; Purification.

Grants and funding

This research was supported by the National Key Research and Development Program of China (No. 2018YFC1602902 & 2016YFD0500701) and High-Level Talent Internationalization Training Project of Henan Province. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.