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Despite high levels of vaccination, currently available commercial vaccines are limited by safety concerns, moderate efficacy, and required high doses. The development of new vaccines is therefore essential. Vaccine efforts should focus on optimizing antigen presentation to enhance immune responses. Here, we describe a simple herringbone\u2010dimer strategy for efficient vaccine design, using the classical swine fever virus E2 expressed in a rice endosperm as an example. The expression of rE2 protein was identified, with the rE2 antigen accumulating to 480\u2009mg\/kg. Immunological assays in mice, rabbits, and pigs showed high antigenicity of rE2. Two immunizations with 284\u2009ng of the rE2 vaccine or one shot with 5.12\u2009\u03bcg provided effective protection in pigs without interference from pre\u2010existing antibodies. Crystal structure and small\u2010angle X\u2010ray scattering results confirmed the stable herringbone dimeric conformation, which had two fully exposed duplex receptor binding domains. Our results demonstrated that rice endosperm is a promising platform for precise vaccine design, and this strategy can be universally applied to other Flaviviridae virus vaccines.<\/jats:p>","DOI":"10.1111\/pbi.14152","type":"journal-article","created":{"date-parts":[[2023,8,12]],"date-time":"2023-08-12T14:03:52Z","timestamp":1691849032000},"page":"2546-2559","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Rice\u2010produced classical swine fever virus glycoprotein\n                    <scp>E2<\/scp>\n                    with herringbone\u2010dimer design to enhance immune responses"],"prefix":"10.1111","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3982-5416","authenticated-orcid":false,"given":"Qianru","family":"Xu","sequence":"first","affiliation":[{"name":"Henan Provincial Key Laboratory of Animal Immunology Henan Academy of Agricultural Sciences  Zhengzhou China"},{"name":"International Joint Research Center of National Animal Immunology, College of Veterinary Medicine Henan Agriculture University  Zhengzhou China"},{"name":"School of Basic Medical Sciences Henan University  Kaifeng China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5258-9158","authenticated-orcid":false,"given":"Fanshu","family":"Ma","sequence":"additional","affiliation":[{"name":"International Joint Research Center of National Animal Immunology, College of Veterinary Medicine Henan Agriculture University  Zhengzhou China"},{"name":"CAS Key Laboratory of Nano\u2010Bio Interface, Suzhou Institute of Nano\u2010Tech and Nano\u2010Bionics Chinese Academy of Sciences  Suzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daichang","family":"Yang","sequence":"additional","affiliation":[{"name":"College of Life Science Wuhan University  Wuhan China"},{"name":"Wuhan Healthgen Biotechnology Corp.  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