{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T21:11:20Z","timestamp":1790716280081,"version":"4.1.0"},"reference-count":39,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2022,9,14]],"date-time":"2022-09-14T00:00:00Z","timestamp":1663113600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["U21A20260"],"award-info":[{"award-number":["U21A20260"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["212300410359"],"award-info":[{"award-number":["212300410359"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2022ZC65"],"award-info":[{"award-number":["2022ZC65"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["BBS\/OS\/NW\/000007"],"award-info":[{"award-number":["BBS\/OS\/NW\/000007"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006407","name":"Natural Science Foundation of Henan Province","doi-asserted-by":"publisher","award":["U21A20260"],"award-info":[{"award-number":["U21A20260"]}],"id":[{"id":"10.13039\/501100006407","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006407","name":"Natural Science Foundation of Henan Province","doi-asserted-by":"publisher","award":["212300410359"],"award-info":[{"award-number":["212300410359"]}],"id":[{"id":"10.13039\/501100006407","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006407","name":"Natural Science Foundation of Henan Province","doi-asserted-by":"publisher","award":["2022ZC65"],"award-info":[{"award-number":["2022ZC65"]}],"id":[{"id":"10.13039\/501100006407","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006407","name":"Natural Science Foundation of Henan Province","doi-asserted-by":"publisher","award":["BBS\/OS\/NW\/000007"],"award-info":[{"award-number":["BBS\/OS\/NW\/000007"]}],"id":[{"id":"10.13039\/501100006407","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Independent Innovation Project of Henan Academy of Agricultural Sciences","award":["U21A20260"],"award-info":[{"award-number":["U21A20260"]}]},{"name":"Independent Innovation Project of Henan Academy of Agricultural Sciences","award":["212300410359"],"award-info":[{"award-number":["212300410359"]}]},{"name":"Independent Innovation Project of Henan Academy of Agricultural Sciences","award":["2022ZC65"],"award-info":[{"award-number":["2022ZC65"]}]},{"name":"Independent Innovation Project of Henan Academy of Agricultural Sciences","award":["BBS\/OS\/NW\/000007"],"award-info":[{"award-number":["BBS\/OS\/NW\/000007"]}]},{"name":"BBSRC Newton Fund Joint Centre Awards on \u201cUK-China Centre of Excellence for Research on Avian Diseases\u201d","award":["U21A20260"],"award-info":[{"award-number":["U21A20260"]}]},{"name":"BBSRC Newton Fund Joint Centre Awards on \u201cUK-China Centre of Excellence for Research on Avian Diseases\u201d","award":["212300410359"],"award-info":[{"award-number":["212300410359"]}]},{"name":"BBSRC Newton Fund Joint Centre Awards on \u201cUK-China Centre of Excellence for Research on Avian Diseases\u201d","award":["2022ZC65"],"award-info":[{"award-number":["2022ZC65"]}]},{"name":"BBSRC Newton Fund Joint Centre Awards on \u201cUK-China Centre of Excellence for Research on Avian Diseases\u201d","award":["BBS\/OS\/NW\/000007"],"award-info":[{"award-number":["BBS\/OS\/NW\/000007"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Marek\u2019s disease virus (MDV) is an important oncogenic \u03b1-herpesvirus that induces Marek\u2019s disease (MD), characterized by severe immunosuppression and rapid-onset T-cell lymphomas in its natural chicken hosts. Historically, MD is regarded as an ideal biomedical model for studying virally induced cancers. Monoclonal antibodies (mAbs) against viral or host antigenic epitopes are crucial for virology research, especially in the exploration of gene functions, clinical therapy, and the development of diagnostic reagents. Utilizing the CRISPR\/Cas9-based gene-editing technology, we produced a pp38-deleted MDV-1 mutant\u2014GX0101\u0394pp38\u2014and used it for the rapid screening and identification of pp38-specific mAbs from a pool of MDV-specific antibodies from 34 hybridomas. The cross-staining of parental and mutated MDV plaques with hybridoma supernatants was first performed by immunofluorescence assay (IFA). Four monoclonal hybridomas\u2014namely, 4F9, 31G7, 34F2, and 35G9\u2014were demonstrated to secrete specific antibodies against MDV-1\u2019s pp38 protein, which was further confirmed by IFA staining and confocal analysis. Further experiments using Western blotting, immunoprecipitation (IP), liquid chromatography\u2013tandem mass spectrometry (LC\u2013MS\/MS), and immunohistochemistry (IHC) analysis demonstrated that the pp38-specific mAb 31G7 has high specificity and wide application potential for further research in MD biology. To the best of our knowledge, this is the first demonstration of the use of CRISPR\/Cas9-based gene-editing technology for efficient screening and identification of mAbs against a specific viral protein, and provides a meaningful reference for the future production of antibodies against other viruses\u2014especially for large DNA viruses such as herpesviruses.<\/jats:p>","DOI":"10.3390\/v14092045","type":"journal-article","created":{"date-parts":[[2022,9,15]],"date-time":"2022-09-15T01:48:10Z","timestamp":1663206490000},"page":"2045","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["A New Strategy for Efficient Screening and Identification of Monoclonal Antibodies against Oncogenic Avian Herpesvirus Utilizing CRISPR\/Cas9-Based Gene-Editing Technology"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2784-4169","authenticated-orcid":false,"given":"Man","family":"Teng","sequence":"first","affiliation":[{"name":"Key Laboratory of Animal Immunology, Ministry of Agriculture and Rural Affairs of China & Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China"},{"name":"UK-China Centre of Excellence for Research on Avian Diseases, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zi-Yu","family":"Zhou","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Immunology, Ministry of Agriculture and Rural Affairs of China & Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China"},{"name":"UK-China Centre of Excellence for Research on Avian Diseases, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China"},{"name":"College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongxiu","family":"Yao","sequence":"additional","affiliation":[{"name":"The Pirbright Institute & UK-China Centre of Excellence for Research on Avian Diseases, Pirbright, Ash Road, Guildford GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8255-516X","authenticated-orcid":false,"given":"Venugopal","family":"Nair","sequence":"additional","affiliation":[{"name":"The Pirbright Institute & UK-China Centre of Excellence for Research on Avian Diseases, Pirbright, Ash Road, Guildford GU24 0NF, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gai-Ping","family":"Zhang","sequence":"additional","affiliation":[{"name":"International Joint Research Center of National Animal Immunology & College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou University, Yangzhou 225009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7351-8180","authenticated-orcid":false,"given":"Jun","family":"Luo","sequence":"additional","affiliation":[{"name":"Key Laboratory of Animal Immunology, Ministry of Agriculture and Rural Affairs of China & Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China"},{"name":"UK-China Centre of Excellence for Research on Avian Diseases, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China"},{"name":"College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1080\/03079457.2011.646238","article-title":"The long view: 40 years of Marek\u2019s disease research and Avian Pathology","volume":"41","author":"Biggs","year":"2012","journal-title":"Avian Pathol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1637\/11525-110216-Reg.1","article-title":"Industry-Wide Surveillance of Marek\u2019s Disease Virus on Commercial Poultry Farms","volume":"61","author":"Kennedy","year":"2017","journal-title":"Avian Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"001673","DOI":"10.1099\/jgv.0.001673","article-title":"ICTV Virus Taxonomy Profile: Herpesviridae 2021","volume":"102","author":"Gatherer","year":"2021","journal-title":"J. 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