Academic Journal
Development and epitope mapping of monoclonal antibodies against group a porcine rotavirus VP6 protein.
| Τίτλος: | Development and epitope mapping of monoclonal antibodies against group a porcine rotavirus VP6 protein. |
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| Συγγραφείς: | 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. |
| Πηγή: | International journal of biological macromolecules [Int J Biol Macromol] 2026 Jun; Vol. 366, pp. 152548. Date of Electronic Publication: 2026 May 14. |
| Τύπος έκδοσης: | Journal Article |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | 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. |
| Ιατρικοί όροι (MeSH): | 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 |
| Περίληψη: | 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 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1879-0003 |
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| DOI: | 10.1016/j.ijbiomac.2026.152548 |