Generation and characterization of a spike glycoprotein domain a-specific neutralizing single-chain variable fragment against porcine epidemic diarrhea virus
Journal
Vaccines
Journal Volume
9
Journal Issue
8
Date Issued
2021
Author(s)
Abstract
The emergence of the genotype (G) 2 and re-emergence of the G1 porcine epidemic diarrhea virus (PEDV) has caused severe economic impacts in the past decade. Developments of efficient vaccines against new variants of PEDV have been challenging, not least because of the difficulties in eliciting mucosal and lactogenic immunity. A single-chain fragment variable (scFv) capable of efficient antigen recognition is an alternative to vaccination and treatment of a viral infection. In the present study, the variable regions of the light chain and the heavy chain of a G2b PEDV spike domain A (S1A)-specific neutralizing monoclonal antibody (mAb) were sequenced, constructed with a (G4S) x3 linker, and produced by a mammalian protein expression system. Our results demonstrated that the PEDV S1A domain scFv was able to bind to S proteins of both G1 and G2b PEDVs. Nevertheless, the scFv was only capable of neutralizing the homologous G2b PEDV but not the G1 PEDV. The binding ability of the G2b-specific neutralizing scFv was not able to predict the neutralizing ability toward heterologous PEDV. The anti-PEDV S1A scFv presented herein serves as a potential therapeutic candidate against the virulent G2b PEDV. ? 2021 by the authors.
Subjects
Neutralizing antibody
Porcine epidemic diarrhea virus
Single-chain variable fragment (scFv)
agar
brilliant blue g
buffer
expi293
expifectamine
fluorescein isothiocyanate
horseradish peroxidase
imidazole
immunoglobulin heavy chain
immunoglobulin kappa chain
immunoglobulin lambda chain
kpl
peroxidase
phosphate buffered saline
r250
reagent
RNA directed DNA polymerase
single chain fragment variable antibody
smartscribe
sodium azide
sodium chloride
tissue plasminogen activator
virus spike protein
animal cell
antigen recognition
Article
controlled study
DNA sequence
epitope mapping
fast protein liquid chromatography
gene expression system
genetic transfection
hybridoma cell line
immobilized metal affinity chromatography
immune response
immunocytochemistry
immunofluorescence assay
immunoprecipitation
live cell imaging
nonhuman
polyacrylamide gel electrophoresis
protein analysis
protein domain
protein expression
reverse transcription polymerase chain reaction
RNA extraction
size exclusion chromatography
spike glycoprotein domain a
Western blotting
SDGs
Type
journal article