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

Abstract

BackgroundAffinity peptides, as a core part of affinity chromatography, play an important role in the purification of target molecules.MethodsHere 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.ResultsOur data showed that the peptide L11- DYWWQSWE has the smallest KD = 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.ConclusionBased 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.

PCV2 Triggers PK-15 Cell Apoptosis Through the PLC–IP3R–Ca2+ Signaling Pathway

“…After the virus enters the host cell, it replicates itself with the host’s help and threat ( Hao et al, 2019 ). Viruses seek advantages, avoid disadvantages, and also affect the internal environment of host cells.…”

New discovery of high-affinity SARS-CoV-2 spike S2 protein binding peptide selected by PhIP-Seq

“…Since S2 subunit performs an important role in viral infusion and entry, it is also more conserved to be the target site for anti-viral inhibitor development ( Ng and Mohd-Ismail, 2021 ). Our data showed that Spep-1 had a high affinity to SARS-CoV-2 spike trimer and monomer protein, with the K D values of 1.70 and 2.14 nmol/L (Spep-1 as the ligand in SPR analysis), which are quite high among the reported peptides ( Mendoza-Figueroa et al, 2018 ; Hao et al, 2019 ; Pomplun et al, 2021 ; Rosestedt et al, 2021 ), suggesting that Spep-1 may have good application prospects, not only limited to SARS-CoV-2 spike antigen detection. In future studies, with rational modifications, Spep-1 might be used for studying the spatial and temporal distribution of S2 subunit during virus infusion, in vivo virus tracing, and drug delivery in biosafety laboratories.…”

Selection and Characterization of a Single-Chain Variable Fragment against Porcine Circovirus Type 2 Capsid and Impedimetric Immunosensor Development

“…21 Yang (K D = 49.4 nM) 43 observed kinetics binding by using sdAb. Hao (K D = 103 nM) 39 and Zheng (K D = 98.5 nM) 38 observed kinetics binding using peptides against PCV2. However, no report has evaluated the binding affinity of the scFv antibody to PCV2 using SPR analysis.…”

“…Phage display screening has previously been used for selecting the specific binder to the PCV2 capsid, but different libraries exist such as peptide libraries − and Bactrian camel sdAb library. − This study used human scFv library. The single-domain antibody (sdAb) has smaller fragments with size 15 kDa consisting of either VH or VL domains, while scFv consists of both VH and VL with a size of 28 kDa.…”