Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2
ABSTRACT: Since 2015, outbreaks of a disease causing severe visceral gout in goslings had resulted in substantial economic losses to the goose farming industry in China. Subsequently, the disease, characterized by extensive visceral urate deposition and renal swelling, was determined to be caused by...
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Elsevier
2025-08-01
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| Series: | Poultry Science |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579125006005 |
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| author | Wei Li Linhua Xu Hantai Tan Tao Wang Zhen Wu Yu He Mingshu Wang Renyong Jia Dekang Zhu Mafeng Liu Xinxin Zhao Qiao Yang Ying Wu Shaqiu Zhang Juan Huang Xumin Ou Di Sun Bin Tian Anchun Cheng Shun Chen |
| author_facet | Wei Li Linhua Xu Hantai Tan Tao Wang Zhen Wu Yu He Mingshu Wang Renyong Jia Dekang Zhu Mafeng Liu Xinxin Zhao Qiao Yang Ying Wu Shaqiu Zhang Juan Huang Xumin Ou Di Sun Bin Tian Anchun Cheng Shun Chen |
| author_sort | Wei Li |
| collection | DOAJ |
| description | ABSTRACT: Since 2015, outbreaks of a disease causing severe visceral gout in goslings had resulted in substantial economic losses to the goose farming industry in China. Subsequently, the disease, characterized by extensive visceral urate deposition and renal swelling, was determined to be caused by a novel astrovirus, designated as goose astrovirus (GAstV). The capsid protein (Cap) of GAstV, encoded by ORF2, is the sole structural protein of the virus and holds potential for developing therapeutic antibodies and diagnostic tools. Based on genetic divergence in the ORF2 gene, GAstV is classified into two serotypes: GAstV-1 and GAstV-2. Despite the critical role of the GAstV Cap in viral pathogenesis, research on generating and characterizing monoclonal antibodies (mAb) against this antigen remains scarce. In this study, six mAbs (1B3, 1B4, 1B6, 1D4, 1E2, 1F1) specifically recognizing GAstV Cap were screened using Western blotting (WB), indirect immunofluorescence assay (IFA), and indirect enzyme-linked immunosorbent assay (ELISA). For epitope mapping, sequential truncations of the GAstV Cap protein fused to glutathione S-transferase (GST) were generated using bacterial expression systems. Ultimately, antigenicity analysis of the prokaryotically expressed, GST-tagged Cap truncations via indirect ELISA and WB delineated two minimal linear epitopes: epitope 630TDPEED635, recognized by mAbs 1B3, 1B4, 1B6, 1D4, and 1E2, and epitope 3DRAVAPREK11, recognized by mAb 1F1. Amino acid sequence alignment revealed that the sequences of epitopes 630TDPEED635 and 3DRAVAPREK11 are highly conserved in GAstV-2 but exhibit significant divergence in the GAstV-1 serotype. This study provides essential tools for both fundamental research and diagnostics of GAstV-2. |
| format | Article |
| id | doaj-art-d5dde5fb47864524bbbb897279d25626 |
| institution | DOAJ |
| issn | 0032-5791 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Elsevier |
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| series | Poultry Science |
| spelling | doaj-art-d5dde5fb47864524bbbb897279d256262025-08-20T02:50:09ZengElsevierPoultry Science0032-57912025-08-01104810535710.1016/j.psj.2025.105357Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2Wei Li0Linhua Xu1Hantai Tan2Tao Wang3Zhen Wu4Yu He5Mingshu Wang6Renyong Jia7Dekang Zhu8Mafeng Liu9Xinxin Zhao10Qiao Yang11Ying Wu12Shaqiu Zhang13Juan Huang14Xumin Ou15Di Sun16Bin Tian17Anchun Cheng18Shun Chen19Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,; Key Laboratory of Agricultural Bioinformatics of Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan 611130, China.Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,; Institute of Veterinary Immunology and Green Drugs, Veterinary Department in College of Animal Science, State Key Laboratory of Green Pesticide, Guizhou University, Guiyang 550025, China.Institute of Veterinary Medicine and Immunology, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, China;; Engineering Research Center of Southwest Animal Disease Prevention and Control Technology for Ministry of Education of the People's Republic of China,; Key Laboratory of Agricultural Bioinformatics of Ministry of Education, Sichuan Agricultural University, Chengdu, Sichuan 611130, China.; Correspondence: (S.C.); Tel.: +86-028-86291482ABSTRACT: Since 2015, outbreaks of a disease causing severe visceral gout in goslings had resulted in substantial economic losses to the goose farming industry in China. Subsequently, the disease, characterized by extensive visceral urate deposition and renal swelling, was determined to be caused by a novel astrovirus, designated as goose astrovirus (GAstV). The capsid protein (Cap) of GAstV, encoded by ORF2, is the sole structural protein of the virus and holds potential for developing therapeutic antibodies and diagnostic tools. Based on genetic divergence in the ORF2 gene, GAstV is classified into two serotypes: GAstV-1 and GAstV-2. Despite the critical role of the GAstV Cap in viral pathogenesis, research on generating and characterizing monoclonal antibodies (mAb) against this antigen remains scarce. In this study, six mAbs (1B3, 1B4, 1B6, 1D4, 1E2, 1F1) specifically recognizing GAstV Cap were screened using Western blotting (WB), indirect immunofluorescence assay (IFA), and indirect enzyme-linked immunosorbent assay (ELISA). For epitope mapping, sequential truncations of the GAstV Cap protein fused to glutathione S-transferase (GST) were generated using bacterial expression systems. Ultimately, antigenicity analysis of the prokaryotically expressed, GST-tagged Cap truncations via indirect ELISA and WB delineated two minimal linear epitopes: epitope 630TDPEED635, recognized by mAbs 1B3, 1B4, 1B6, 1D4, and 1E2, and epitope 3DRAVAPREK11, recognized by mAb 1F1. Amino acid sequence alignment revealed that the sequences of epitopes 630TDPEED635 and 3DRAVAPREK11 are highly conserved in GAstV-2 but exhibit significant divergence in the GAstV-1 serotype. This study provides essential tools for both fundamental research and diagnostics of GAstV-2.http://www.sciencedirect.com/science/article/pii/S0032579125006005Goose astrovirusCapsid proteinMonoclonal antibodyLinear epitope |
| spellingShingle | Wei Li Linhua Xu Hantai Tan Tao Wang Zhen Wu Yu He Mingshu Wang Renyong Jia Dekang Zhu Mafeng Liu Xinxin Zhao Qiao Yang Ying Wu Shaqiu Zhang Juan Huang Xumin Ou Di Sun Bin Tian Anchun Cheng Shun Chen Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2 Poultry Science Goose astrovirus Capsid protein Monoclonal antibody Linear epitope |
| title | Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2 |
| title_full | Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2 |
| title_fullStr | Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2 |
| title_full_unstemmed | Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2 |
| title_short | Identification and Characterization of Linear Epitopes of Monoclonal Antibodies Against the Capsid Protein of Goose Astrovirus Genotype 2 |
| title_sort | identification and characterization of linear epitopes of monoclonal antibodies against the capsid protein of goose astrovirus genotype 2 |
| topic | Goose astrovirus Capsid protein Monoclonal antibody Linear epitope |
| url | http://www.sciencedirect.com/science/article/pii/S0032579125006005 |
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