Academic Journal
View in EDS

Development and epitope mapping of monoclonal antibodies against group a porcine rotavirus VP6 protein.

Bibliographic Details
Title: Development and epitope mapping of monoclonal antibodies against group a porcine rotavirus VP6 protein.
Authors: Hu Y; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Wu H; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Xiao Y; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Gao X; China Animal Health and Epidemiology Center, Qingdao, 266032, PR China., Sun R; Qingdao Lijian Bio-tech Co., Ltd., Qingdao, Shandong, 266041, PR China., Xin W; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Chen Z; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Wu X; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Ding Z; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Song D; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China., Wu H; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China. Electronic address: huanswu@jxau.edu.cn.
Source: International journal of biological macromolecules [Int J Biol Macromol] 2026 Jun; Vol. 366, pp. 152548. Date of Electronic Publication: 2026 May 14.
Publication Type: Journal Article
Language: English
Journal Info: Publisher: Elsevier Country of Publication: Netherlands NLM ID: 7909578 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0003 (Electronic) Linking ISSN: 01418130 NLM ISO Abbreviation: Int J Biol Macromol Subsets: MEDLINE
Imprint Name(s): Publication: Amsterdam : Elsevier
Original Publication: Guildford, Eng., IPC Science and Technology Press.
MeSH Terms: Antibodies, Monoclonal*/immunology , Capsid Proteins*/immunology , Capsid Proteins*/chemistry , Capsid Proteins*/genetics , Antigens, Viral*/immunology , Antigens, Viral*/chemistry , Antigens, Viral*/genetics , Rotavirus*/immunology , Rotavirus*/genetics , Epitope Mapping*/methods, Epitopes, B-Lymphocyte/immunology ; Epitopes, B-Lymphocyte/chemistry ; Antibodies, Viral/immunology ; Recombinant Proteins/immunology ; Animals ; Swine ; Amino Acid Sequence ; Molecular Docking Simulation
Abstract: Porcine rotavirus (PoRV) is a major causative agent of viral diarrhea in piglets. Its VP6 protein, the most abundant structural component of the viral particle, is highly conserved and immunogenic, making it an ideal target for developing broad-spectrum diagnostics and investigating cross-protective immunity. In this study, the PoRV VP6 recombinant protein was successfully produced and purified using a prokaryotic expression system. The recombinant VP6 protein was successfully expressed in E. coli and purified, with its purity assessed by RP-HPLC (61.43%). Four hybridoma cell lines (1D9, 2E7, 3G11, and 5E3) stably secreting monoclonal antibodies (mAbs) against VP6 were established via hybridoma technology. These mAbs exhibited strong reactivity and high specificity, as confirmed by indirect ELISA, Western blot, and indirect immunofluorescence assays. Furthermore, employing a strategy of serial truncation and alanine scanning, we identified the four linear B-cell epitopes on the VP6 protein: 43NDFQTGG49, 51GNLPIRNWTF60, 208EHVVPLR214, and 237INSAD241. Conservation analysis revealed that the epitopes recognized by mAbs 1D9 and 3G11 were highly conserved across geographically distinct strains, whereas those recognized by mAbs 5E3 and 2E7 displayed notable region-specific characteristics. Molecular docking simulations demonstrated that the mAbs formed stable complexes with their corresponding epitopes primarily through hydrogen bonding, electrostatic interactions, and hydrophobic forces, with mAb 5E3 exhibiting the highest affinity (Docking Score: -208.4). The mAbs obtained in this study, together with the precise mapping of their epitopes, provide crucial tools and a foundation for advancing PoRV serological detection methods and studying the antigenic structure of the VP6 protein.
(Copyright © 2026 Elsevier B.V. All rights reserved.)
Competing Interests: Declaration of competing interest The authors report no conflicts of interest.
Contributed Indexing: Keywords: Epitope mapping; Monoclonal antibodies; Porcine rotavirus; VP6
Substance Nomenclature: 0 (VP6 protein, Rotavirus)
0 (Antibodies, Monoclonal)
0 (Capsid Proteins)
0 (Antigens, Viral)
0 (Epitopes, B-Lymphocyte)
0 (Antibodies, Viral)
0 (Recombinant Proteins)
Entry Date(s): Date Created: 20260515 Date Completed: 20260716 Latest Revision: 20260716
Update Code: 20260717
DOI: 10.1016/j.ijbiomac.2026.152548
PMID: 42140294
Database: MEDLINE
FullText Text:
  Availability: 0
CustomLinks:
  – Url: https://www.doi.org/10.1016/j.ijbiomac.2026.152548?
    Name: ScienceDirect (all content) (s7799221)
    Category: fullText
    Text: View record from ScienceDirect
    MouseOverText: View record from ScienceDirect
  – Url: https://resolver.ebsco.com/c/fiv2js/result?sid=EBSCO:cmedm&genre=article&issn=18790003&ISBN=&volume=366&issue=&date=20260601&spage=152548&pages=152548&title=International journal of biological macromolecules&atitle=Development%20and%20epitope%20mapping%20of%20monoclonal%20antibodies%20against%20group%20a%20porcine%20rotavirus%20VP6%20protein.&aulast=Hu%20Y&id=DOI:10.1016/j.ijbiomac.2026.152548
    Name: Full Text Finder (for New FTF UI) (ns324271)
    Category: fullText
    Text: Full Text Finder
    MouseOverText: Full Text Finder
Header DbId: cmedm
DbLabel: MEDLINE
An: 42140294
AccessLevel: 3
PubType: Academic Journal
PubTypeId: academicJournal
PreciseRelevancyScore: 0
IllustrationInfo
Items – Name: Title
  Label: Title
  Group: Ti
  Data: Development and epitope mapping of monoclonal antibodies against group a porcine rotavirus VP6 protein.
– Name: Author
  Label: Authors
  Group: Au
  Data: <searchLink fieldCode="AU" term="%22Hu+Y%22">Hu Y</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Wu+H%22">Wu H</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Xiao+Y%22">Xiao Y</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Gao+X%22">Gao X</searchLink>; China Animal Health and Epidemiology Center, Qingdao, 266032, PR China.<br /><searchLink fieldCode="AU" term="%22Sun+R%22">Sun R</searchLink>; Qingdao Lijian Bio-tech Co., Ltd., Qingdao, Shandong, 266041, PR China.<br /><searchLink fieldCode="AU" term="%22Xin+W%22">Xin W</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Chen+Z%22">Chen Z</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Wu+X%22">Wu X</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Ding+Z%22">Ding Z</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Song+D%22">Song D</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China.<br /><searchLink fieldCode="AU" term="%22Wu+H%22">Wu H</searchLink>; Department of Veterinary Preventive Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China; Jiangxi Provincial Key Laboratory for Animal Health, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, 330045, PR China. Electronic address: huanswu@jxau.edu.cn.
– Name: TitleSource
  Label: Source
  Group: Src
  Data: <searchLink fieldCode="JN" term="%227909578%22">International journal of biological macromolecules</searchLink> [Int J Biol Macromol] 2026 Jun; Vol. 366, pp. 152548. <i>Date of Electronic Publication: </i>2026 May 14.
– Name: TypePub
  Label: Publication Type
  Group: TypPub
  Data: Journal Article
– Name: Language
  Label: Language
  Group: Lang
  Data: English
– Name: TitleSource
  Label: Journal Info
  Group: Src
  Data: <i>Publisher: </i><searchLink fieldCode="PB" term="%22Elsevier%22">Elsevier </searchLink><i>Country of Publication: </i>Netherlands <i>NLM ID: </i>7909578 <i>Publication Model: </i>Print-Electronic <i>Cited Medium: </i>Internet <i>ISSN: </i>1879-0003 (Electronic) <i>Linking ISSN: </i><searchLink fieldCode="IS" term="%2201418130%22">01418130 </searchLink><i>NLM ISO Abbreviation: </i>Int J Biol Macromol <i>Subsets: </i>MEDLINE
– Name: PublisherInfo
  Label: Imprint Name(s)
  Group: PubInfo
  Data: <i>Publication</i>: Amsterdam : Elsevier<br /><i>Original Publication</i>: Guildford, Eng., IPC Science and Technology Press.
– Name: SubjectMESH
  Label: MeSH Terms
  Group: Su
  Data: <searchLink fieldCode="MM" term="%22Antibodies%2C+Monoclonal%22">Antibodies, Monoclonal*</searchLink>/<searchLink fieldCode="MM" term="%22Antibodies%2C+Monoclonal+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Capsid+Proteins%22">Capsid Proteins*</searchLink>/<searchLink fieldCode="MM" term="%22Capsid+Proteins+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Capsid+Proteins%22">Capsid Proteins*</searchLink>/<searchLink fieldCode="MM" term="%22Capsid+Proteins+chemistry%22">chemistry</searchLink> <br /><searchLink fieldCode="MM" term="%22Capsid+Proteins%22">Capsid Proteins*</searchLink>/<searchLink fieldCode="MM" term="%22Capsid+Proteins+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Antigens%2C+Viral%22">Antigens, Viral*</searchLink>/<searchLink fieldCode="MM" term="%22Antigens%2C+Viral+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Antigens%2C+Viral%22">Antigens, Viral*</searchLink>/<searchLink fieldCode="MM" term="%22Antigens%2C+Viral+chemistry%22">chemistry</searchLink> <br /><searchLink fieldCode="MM" term="%22Antigens%2C+Viral%22">Antigens, Viral*</searchLink>/<searchLink fieldCode="MM" term="%22Antigens%2C+Viral+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Rotavirus%22">Rotavirus*</searchLink>/<searchLink fieldCode="MM" term="%22Rotavirus+immunology%22">immunology</searchLink> <br /><searchLink fieldCode="MM" term="%22Rotavirus%22">Rotavirus*</searchLink>/<searchLink fieldCode="MM" term="%22Rotavirus+genetics%22">genetics</searchLink> <br /><searchLink fieldCode="MM" term="%22Epitope+Mapping%22">Epitope Mapping*</searchLink>/<searchLink fieldCode="MM" term="%22Epitope+Mapping+methods%22">methods</searchLink><br /><searchLink fieldCode="MH" term="%22Epitopes%2C+B-Lymphocyte%22">Epitopes, B-Lymphocyte</searchLink>/<searchLink fieldCode="MH" term="%22Epitopes%2C+B-Lymphocyte+immunology%22">immunology</searchLink> ; <searchLink fieldCode="MH" term="%22Epitopes%2C+B-Lymphocyte%22">Epitopes, B-Lymphocyte</searchLink>/<searchLink fieldCode="MH" term="%22Epitopes%2C+B-Lymphocyte+chemistry%22">chemistry</searchLink> ; <searchLink fieldCode="MH" term="%22Antibodies%2C+Viral%22">Antibodies, Viral</searchLink>/<searchLink fieldCode="MH" term="%22Antibodies%2C+Viral+immunology%22">immunology</searchLink> ; <searchLink fieldCode="MH" term="%22Recombinant+Proteins%22">Recombinant Proteins</searchLink>/<searchLink fieldCode="MH" term="%22Recombinant+Proteins+immunology%22">immunology</searchLink> ; <searchLink fieldCode="MH" term="%22Animals%22">Animals</searchLink> ; <searchLink fieldCode="MH" term="%22Swine%22">Swine</searchLink> ; <searchLink fieldCode="MH" term="%22Amino+Acid+Sequence%22">Amino Acid Sequence</searchLink> ; <searchLink fieldCode="MH" term="%22Molecular+Docking+Simulation%22">Molecular Docking Simulation</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: Porcine rotavirus (PoRV) is a major causative agent of viral diarrhea in piglets. Its VP6 protein, the most abundant structural component of the viral particle, is highly conserved and immunogenic, making it an ideal target for developing broad-spectrum diagnostics and investigating cross-protective immunity. In this study, the PoRV VP6 recombinant protein was successfully produced and purified using a prokaryotic expression system. The recombinant VP6 protein was successfully expressed in E. coli and purified, with its purity assessed by RP-HPLC (61.43%). Four hybridoma cell lines (1D9, 2E7, 3G11, and 5E3) stably secreting monoclonal antibodies (mAbs) against VP6 were established via hybridoma technology. These mAbs exhibited strong reactivity and high specificity, as confirmed by indirect ELISA, Western blot, and indirect immunofluorescence assays. Furthermore, employing a strategy of serial truncation and alanine scanning, we identified the four linear B-cell epitopes on the VP6 protein: <superscript>43</superscript>NDFQTGG<superscript>49</superscript>, <superscript>51</superscript>GNLPIRNWTF<superscript>60</superscript>, <superscript>208</superscript>EHVVPLR<superscript>214</superscript>, and <superscript>237</superscript>INSAD<superscript>241</superscript>. Conservation analysis revealed that the epitopes recognized by mAbs 1D9 and 3G11 were highly conserved across geographically distinct strains, whereas those recognized by mAbs 5E3 and 2E7 displayed notable region-specific characteristics. Molecular docking simulations demonstrated that the mAbs formed stable complexes with their corresponding epitopes primarily through hydrogen bonding, electrostatic interactions, and hydrophobic forces, with mAb 5E3 exhibiting the highest affinity (Docking Score: -208.4). The mAbs obtained in this study, together with the precise mapping of their epitopes, provide crucial tools and a foundation for advancing PoRV serological detection methods and studying the antigenic structure of the VP6 protein.<br /> (Copyright © 2026 Elsevier B.V. All rights reserved.)
– Name: Abstract
  Label: Competing Interests
  Group: Ab
  Data: Declaration of competing interest The authors report no conflicts of interest.
– Name: SubjectMinor
  Label: Contributed Indexing
  Group:
  Data: <i>Keywords: </i>Epitope mapping; Monoclonal antibodies; Porcine rotavirus; VP6
– Name: NumberCAS
  Label: Substance Nomenclature
  Group: ID
  Data: 0 (VP6 protein, Rotavirus)<br />0 (Antibodies, Monoclonal)<br />0 (Capsid Proteins)<br />0 (Antigens, Viral)<br />0 (Epitopes, B-Lymphocyte)<br />0 (Antibodies, Viral)<br />0 (Recombinant Proteins)
– Name: DateEntry
  Label: Entry Date(s)
  Group: Date
  Data: <i>Date Created: </i>20260515 <i>Date Completed: </i>20260716 <i>Latest Revision: </i>20260716
– Name: DateUpdate
  Label: Update Code
  Group: Date
  Data: 20260717
– Name: DOI
  Label: DOI
  Group: ID
  Data: 10.1016/j.ijbiomac.2026.152548
– Name: AN
  Label: PMID
  Group: ID
  Data: 42140294
PLink https://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=cmedm&AN=42140294
RecordInfo BibRecord:
  BibEntity:
    Identifiers:
      – Type: doi
        Value: 10.1016/j.ijbiomac.2026.152548
    Languages:
      – Code: eng
        Text: English
    PhysicalDescription:
      Pagination:
        StartPage: 152548
    Subjects:
      – SubjectFull: Epitopes, B-Lymphocyte immunology
        Type: general
      – SubjectFull: Epitopes, B-Lymphocyte chemistry
        Type: general
      – SubjectFull: Antibodies, Viral immunology
        Type: general
      – SubjectFull: Recombinant Proteins immunology
        Type: general
      – SubjectFull: Animals
        Type: general
      – SubjectFull: Swine
        Type: general
      – SubjectFull: Amino Acid Sequence
        Type: general
      – SubjectFull: Molecular Docking Simulation
        Type: general
      – SubjectFull: Antibodies, Monoclonal immunology
        Type: general
      – SubjectFull: Capsid Proteins immunology
        Type: general
      – SubjectFull: Capsid Proteins chemistry
        Type: general
      – SubjectFull: Capsid Proteins genetics
        Type: general
      – SubjectFull: Antigens, Viral immunology
        Type: general
      – SubjectFull: Antigens, Viral chemistry
        Type: general
      – SubjectFull: Antigens, Viral genetics
        Type: general
      – SubjectFull: Rotavirus immunology
        Type: general
      – SubjectFull: Rotavirus genetics
        Type: general
      – SubjectFull: Epitope Mapping methods
        Type: general
    Titles:
      – TitleFull: Development and epitope mapping of monoclonal antibodies against group a porcine rotavirus VP6 protein.
        Type: main
  BibRelationships:
    HasContributorRelationships:
      – PersonEntity:
          Name:
            NameFull: Hu Y
      – PersonEntity:
          Name:
            NameFull: Wu H
      – PersonEntity:
          Name:
            NameFull: Xiao Y
      – PersonEntity:
          Name:
            NameFull: Gao X
      – PersonEntity:
          Name:
            NameFull: Sun R
      – PersonEntity:
          Name:
            NameFull: Xin W
      – PersonEntity:
          Name:
            NameFull: Chen Z
      – PersonEntity:
          Name:
            NameFull: Wu X
      – PersonEntity:
          Name:
            NameFull: Ding Z
      – PersonEntity:
          Name:
            NameFull: Song D
      – PersonEntity:
          Name:
            NameFull: Wu H
    IsPartOfRelationships:
      – BibEntity:
          Dates:
            – D: 01
              M: 06
              Text: 2026 Jun
              Type: published
              Y: 2026
          Identifiers:
            – Type: issn-electronic
              Value: 1879-0003
          Numbering:
            – Type: volume
              Value: 366
          Titles:
            – TitleFull: International journal of biological macromolecules
              Type: main
ResultId 1