Identification of Three Novel Linear B-Cell Epitopes in Non-Structural Protein 3 of Porcine Epidemic Diarrhea Virus Using Monoclonal Antibodies.

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101509722 Publication Model: Electronic Cited Medium: Internet ISSN: 1999-4915 (Electronic) Linking ISSN: 19994915 NLM ISO Abbreviation: Viruses Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI

Porcine epidemic diarrhea virus*/genetics ; Coronavirus Infections* ; Swine Diseases*; Animals ; Swine ; Epitopes, B-Lymphocyte ; Antibodies, Monoclonal ; Blotting, Western ; Amino Acids

Porcine epidemic diarrhea virus (PEDV) is a highly pathogenic swine coronavirus that causes diarrhea and high mortality in piglets, resulting in significant economic losses within the global swine industry. Nonstructural protein 3 (Nsp3) is the largest in coronavirus, playing critical roles in viral replication, such as the processing of polyproteins and the formation of replication-transcription complexes (RTCs). In this study, three monoclonal antibodies (mAbs), 7G4, 5A3, and 2D7, targeting PEDV Nsp3 were successfully generated, and three distinct linear B-cell epitopes were identified within these mAbs by using Western blotting analysis with 24 truncations of Nsp3. The epitope against 7G4 was located on amino acids 31-TISQDLLDVE-40, the epitope against 5A3 was found on amino acids 141-LGIVDDPAMG-150, and the epitope against 2D7 was situated on amino acids 282-FYDAAMAIDG-291. Intriguingly, the epitope 31-TISQDLLDVE-40 recognized by the mAb 7G4 appears to be a critical B-cell linear epitope due to its high antigenic index and exposed location on the surface of Nsp3 protein. In addition, bioinformatics analysis unveiled that these three epitopes were highly conserved in most genotypes of PEDV. These findings present the first characterization of three novel linear B-cell epitopes in the Nsp3 protein of PEDV and provide potential tools of mAbs for identifying host proteins that may facilitate viral infection.

Autophagy. 2020 Oct;16(10):1737-1752. (PMID: 31868081)
iScience. 2023 Apr 21;26(4):106280. (PMID: 36945252)
Antiviral Res. 2016 Nov;135:97-107. (PMID: 27743916)
Virus Res. 2020 May;281:197912. (PMID: 32142743)
Transbound Emerg Dis. 2020 May;67(3):1129-1140. (PMID: 31785090)
J Virol. 2008 Jun;82(11):5279-94. (PMID: 18367524)
Emerg Microbes Infect. 2021 Dec;10(1):178-195. (PMID: 33372854)
Nat Commun. 2023 Mar 28;14(1):1733. (PMID: 36977673)
Nat Commun. 2023 Jul 6;14(1):3999. (PMID: 37414753)
Emerg Infect Dis. 2012 Aug;18(8):1350-3. (PMID: 22840964)
Adv Exp Med Biol. 2016;917:11-22. (PMID: 27236550)
Front Immunol. 2020 Jul 03;11:1581. (PMID: 32719684)
Elife. 2015 Oct 21;4:. (PMID: 26488441)
Sci Rep. 2023 Feb 18;13(1):2890. (PMID: 36801935)
Curr Opin Virol. 2019 Feb;34:39-49. (PMID: 30654269)
PLoS One. 2011 Feb 18;6(2):e17192. (PMID: 21364999)
J Virol. 2012 Apr;86(7):3995-4008. (PMID: 22278237)
J Virol. 2010 Oct;84(19):10276-88. (PMID: 20660183)
Microb Pathog. 2023 Aug;181:106185. (PMID: 37269880)
J Gen Virol. 2013 Jul;94(Pt 7):1554-1567. (PMID: 23596270)
Int J Mol Sci. 2022 Oct 02;23(19):. (PMID: 36232993)
J Biol Chem. 2021 Sep;297(3):101041. (PMID: 34358560)
Int J Mol Sci. 2022 Aug 26;23(17):. (PMID: 36077070)
Science. 2020 Sep 11;369(6509):1395-1398. (PMID: 32763915)
J Virol. 2022 Jan 12;96(1):e0137221. (PMID: 34643430)
J Vet Diagn Invest. 2013 Sep;25(5):649-54. (PMID: 23963154)
Int J Mol Sci. 2022 Dec 31;24(1):. (PMID: 36614163)
Nature. 2022 Jun;606(7915):761-768. (PMID: 35551511)
J Virol. 2023 Jul 27;97(7):e0196422. (PMID: 37358450)
Virol J. 2019 Aug 5;16(1):97. (PMID: 31382996)
Protein Cell. 2014 May;5(5):369-81. (PMID: 24622840)
Cell Host Microbe. 2017 Aug 9;22(2):176-184. (PMID: 28799903)
J Cell Biol. 2023 Jul 3;222(7):. (PMID: 37093123)
Virology. 2023 Feb;579:29-37. (PMID: 36592554)
Vet Microbiol. 2017 Jul;206:45-51. (PMID: 27964998)
Virus Res. 2019 Jan 2;259:18-27. (PMID: 30342075)
Front Vet Sci. 2022 Apr 08;9:820625. (PMID: 35464391)
Virol J. 2020 Apr 3;17(1):46. (PMID: 32245493)
Vet Microbiol. 2019 Mar;230:278-282. (PMID: 30827401)
Nat Commun. 2020 Jul 30;11(1):3810. (PMID: 32733001)
J Gen Virol. 2014 Mar;95(Pt 3):614-626. (PMID: 24362959)
Sci Rep. 2016 Dec 19;6:39010. (PMID: 27991537)
Mol Ther. 2014 Apr;22(4):881-90. (PMID: 24370701)
Appl Microbiol Biotechnol. 2019 May;103(9):3705-3714. (PMID: 30877355)
J Vet Sci. 2023 Jul;24(4):e58. (PMID: 37532301)
PLoS One. 2012;7(2):e30802. (PMID: 22312431)

2021YFD1801102 the National Key Research and Development Program of China

Keywords: B-cell epitopes; Nsp3 protein; monoclonal antibody; porcine epidemic diarrhea virus

0 (Epitopes, B-Lymphocyte)
0 (Antibodies, Monoclonal)
0 (Amino Acids)

Date Created: 20240328 Date Completed: 20240329 Latest Revision: 20240522

20250114

PMC10975687

10.3390/v16030424

38543789