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

文献类型： 外文期刊

作者： Hao, Junfang 1 ; Wang, Fangyu 2 ; Xing, Guangxu 2 ; Liu, Yunchao 2 ; Deng, Ruiguang 2 ; Zhang, Hao 2 ; Cheng, Anchun 1 ;

作者机构： 1.Sichuan Agr Univ, Coll Vet Med, Res Ctr Avian Dis, Chengdu, Sichuan, Peoples R China

2.Henan Acad Agr Sci, Henan Key Lab Anim Immunol, Zhengzhou, Henan, Peoples R China

3.Henan Agr Univ, Coll Life Sci, Zhengzhou, Henan, Peoples R China

关键词： Affinity peptides; PCV2 Cap; Molecular docking virtual screening; Purification

期刊名称：PEERJ （ 影响因子：2.984； 五年影响因子：3.369 ）

ISSN： 2167-8359

年卷期： 2019 年 7 卷

页码：

收录情况： SCI

摘要： 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.

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