Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virus
Abstract Bluetongue virus (BTV) VP7 is a group-specific protein that is highly conserved in different serotypes. In this study, BALB/c mice were immunized with purified recombinant BTV-1 VP7 protein expressed in E. coli. Then six monoclonal antibodies (mAbs), 2A7, 2B2, 2B3, 2D3, 2D7, and 2H2, agains...
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BMC
2025-04-01
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| Series: | Virology Journal |
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| Online Access: | https://doi.org/10.1186/s12985-025-02733-7 |
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| author | Xin-Bing Hu Jing Xu Yu Zhang Fan-Hua Meng Zhan-Cheng Tian Gui-Quan Guan Jian-Xun Luo Hong Yin Jun-Zheng Du |
| author_facet | Xin-Bing Hu Jing Xu Yu Zhang Fan-Hua Meng Zhan-Cheng Tian Gui-Quan Guan Jian-Xun Luo Hong Yin Jun-Zheng Du |
| author_sort | Xin-Bing Hu |
| collection | DOAJ |
| description | Abstract Bluetongue virus (BTV) VP7 is a group-specific protein that is highly conserved in different serotypes. In this study, BALB/c mice were immunized with purified recombinant BTV-1 VP7 protein expressed in E. coli. Then six monoclonal antibodies (mAbs), 2A7, 2B2, 2B3, 2D3, 2D7, and 2H2, against the BTV-1 VP7 protein were produced using hybridoma technology. The reactivity of the mAbs was identified using western blotting, enzyme-linked immunosorbent assay, and immunofluorescence assay. A series of truncated peptides derived from VP7 expressed as glutathione S-transferase fusion proteins were mapped with mAbs by western blotting. The results indicated that 2A7 recognized the epitope 71SAAGINVGPI80, 2B3 recognized 110ARVTGETSTWG120, 2B2 and 2D3 recognized 125PYGFFLETEET135, and 2D7 and 2H2 recognized 332VNPMPGPLTRA342. Amino acid sequence analysis showed that these four epitopes were conserved in 24 typical BTV serotypes. Cross-reaction results showed that mAb 2A7 could recognize the recombinant VP7 protein of BTV-1, African horse sickness virus serotype 1 (AHSV-1), and epidemic hemorrhagic disease virus serotype 1 (EHDV-1). The mAbs 2B2, 2B3, and 2D3 could recognize the recombinant VP7 protein of BTV-1 and EHDV-1, and the mAbs 2D7 and 2H2 specifically recognized the BTV-1 VP7 protein. These specific mAbs and identified B-cell epitopes provided key insights into the structure and function of VP7, while facilitating the development of BTV diagnostics and the design of epitope-based vaccines. |
| format | Article |
| id | doaj-art-021bb036072846e1ba68b5d2441aa33c |
| institution | OA Journals |
| issn | 1743-422X |
| language | English |
| publishDate | 2025-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj-art-021bb036072846e1ba68b5d2441aa33c2025-08-20T01:47:33ZengBMCVirology Journal1743-422X2025-04-0122111010.1186/s12985-025-02733-7Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virusXin-Bing Hu0Jing Xu1Yu Zhang2Fan-Hua Meng3Zhan-Cheng Tian4Gui-Quan Guan5Jian-Xun Luo6Hong Yin7Jun-Zheng Du8State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesState Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural SciencesAbstract Bluetongue virus (BTV) VP7 is a group-specific protein that is highly conserved in different serotypes. In this study, BALB/c mice were immunized with purified recombinant BTV-1 VP7 protein expressed in E. coli. Then six monoclonal antibodies (mAbs), 2A7, 2B2, 2B3, 2D3, 2D7, and 2H2, against the BTV-1 VP7 protein were produced using hybridoma technology. The reactivity of the mAbs was identified using western blotting, enzyme-linked immunosorbent assay, and immunofluorescence assay. A series of truncated peptides derived from VP7 expressed as glutathione S-transferase fusion proteins were mapped with mAbs by western blotting. The results indicated that 2A7 recognized the epitope 71SAAGINVGPI80, 2B3 recognized 110ARVTGETSTWG120, 2B2 and 2D3 recognized 125PYGFFLETEET135, and 2D7 and 2H2 recognized 332VNPMPGPLTRA342. Amino acid sequence analysis showed that these four epitopes were conserved in 24 typical BTV serotypes. Cross-reaction results showed that mAb 2A7 could recognize the recombinant VP7 protein of BTV-1, African horse sickness virus serotype 1 (AHSV-1), and epidemic hemorrhagic disease virus serotype 1 (EHDV-1). The mAbs 2B2, 2B3, and 2D3 could recognize the recombinant VP7 protein of BTV-1 and EHDV-1, and the mAbs 2D7 and 2H2 specifically recognized the BTV-1 VP7 protein. These specific mAbs and identified B-cell epitopes provided key insights into the structure and function of VP7, while facilitating the development of BTV diagnostics and the design of epitope-based vaccines.https://doi.org/10.1186/s12985-025-02733-7Bluetongue virusVP7 proteinMonoclonal antibodiesB-cell epitope mapping |
| spellingShingle | Xin-Bing Hu Jing Xu Yu Zhang Fan-Hua Meng Zhan-Cheng Tian Gui-Quan Guan Jian-Xun Luo Hong Yin Jun-Zheng Du Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virus Virology Journal Bluetongue virus VP7 protein Monoclonal antibodies B-cell epitope mapping |
| title | Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virus |
| title_full | Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virus |
| title_fullStr | Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virus |
| title_full_unstemmed | Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virus |
| title_short | Monoclonal antibody and B-cell epitope mapping of the VP7 protein in bluetongue virus |
| title_sort | monoclonal antibody and b cell epitope mapping of the vp7 protein in bluetongue virus |
| topic | Bluetongue virus VP7 protein Monoclonal antibodies B-cell epitope mapping |
| url | https://doi.org/10.1186/s12985-025-02733-7 |
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