{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T14:59:20Z","timestamp":1780066760839,"version":"3.54.0"},"reference-count":45,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2019,12,1]],"date-time":"2019-12-01T00:00:00Z","timestamp":1575158400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004396","name":"Thailand Research Fund","doi-asserted-by":"publisher","award":["RSA6280006"],"award-info":[{"award-number":["RSA6280006"]}],"id":[{"id":"10.13039\/501100004396","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Enterovirus 71 (EV71) is the main causative agent of severe hand-foot-mouth disease. EV71 affects countries mainly in the Asia-Pacific region, which makes it unattractive for pharmaceutical companies to develop drugs or vaccine to combat EV71 infection. However, development of these drugs and vaccines is vital to protect younger generations. This study aims to develop a specific monoclonal antibody (mAb) to EV71 using a plant platform, which is a cost-effective and scalable production technology. A previous report showed that D5, a murine anti-EV71 mAb, binds to VP1 protein of EV71, potently neutralizes EV71 in vitro, and effectively protects mice against EV71 infection. Herein, plant-produced chimeric D5 (cD5) mAb, variable regions of murine D5 antibody linked with constant regions of human IgG1, was transiently expressed in Nicotiana benthamiana using geminiviral vectors. The antibody was expressed at high levels within six days of infiltration. Plant-produced cD5 retained its in vitro high-affinity binding and neutralizing activity against EV71. Furthermore, a single dose (10 \u00b5g\/g body weight) of plant-produced cD5 mAb offered 100% protection against infection in mice after a lethal EV71 challenge. Therefore, our results showed that plant-produced anti-EV71 mAb is an effective, safe, and affordable therapeutic option against EV71 infection.<\/jats:p>","DOI":"10.3390\/plants8120560","type":"journal-article","created":{"date-parts":[[2019,12,2]],"date-time":"2019-12-02T10:50:45Z","timestamp":1575283845000},"page":"560","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["Plant-Produced Anti-Enterovirus 71 (EV71) Monoclonal Antibody Efficiently Protects Mice Against EV71 Infection"],"prefix":"10.3390","volume":"8","author":[{"given":"Kaewta","family":"Rattanapisit","sequence":"first","affiliation":[{"name":"Research Unit for Plant-Produced Pharmaceuticals, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8658-9009","authenticated-orcid":false,"given":"Zhang","family":"Chao","sequence":"additional","affiliation":[{"name":"Vaccine Research Center, CAS Key Laboratory of Molecular Virology &amp; Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konlavat","family":"Siriwattananon","sequence":"additional","affiliation":[{"name":"Research Unit for Plant-Produced Pharmaceuticals, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhong","family":"Huang","sequence":"additional","affiliation":[{"name":"Vaccine Research Center, CAS Key Laboratory of Molecular Virology &amp; Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai 200031, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Waranyoo","family":"Phoolcharoen","sequence":"additional","affiliation":[{"name":"Research Unit for Plant-Produced Pharmaceuticals and Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,12,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.4269\/ajtmh.18-0099","article-title":"The effect of school closure on hand, foot, and mouth disease transmission in Singapore: A Modeling Approach","volume":"99","author":"Chen","year":"2018","journal-title":"Am. J. Trop. Med. Hyg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ijid.2018.12.004","article-title":"Clinical and aetiological study of hand, foot and mouth disease in southern Vietnam, 2013-2015: Inpatients and outpatients","volume":"80","author":"Hoang","year":"2019","journal-title":"Int. J. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1097","DOI":"10.1016\/S1474-4422(10)70209-X","article-title":"Clinical features, diagnosis, and management of enterovirus 71","volume":"9","author":"Ooi","year":"2010","journal-title":"Lancet Neurol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"936","DOI":"10.1056\/NEJM199909233411302","article-title":"Neurologic complications in children with enterovirus 71 infection","volume":"341","author":"Huang","year":"1999","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1028","DOI":"10.4161\/hv.6.12.12982","article-title":"Optimized development of a candidate strain of inactivated EV71 vaccine and analysis of its immunogenicity in rhesus monkeys","volume":"6","author":"Dong","year":"2010","journal-title":"Hum. Vaccin."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Liu, L., Zhang, Y., Wang, J., Zhao, H., Jiang, L., Che, Y., Shi, H., Li, R., Mo, Z., and Huang, T. (2013). Study of the integrated immune response induced by an inactivated EV71 vaccine. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0054451"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"6269","DOI":"10.1016\/j.vaccine.2011.06.044","article-title":"Immunoprotection elicited by an enterovirus type 71 experimental inactivated vaccine in mice and rhesus monkeys","volume":"29","author":"Dong","year":"2011","journal-title":"Vaccine"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"4829","DOI":"10.1016\/j.vaccine.2011.04.070","article-title":"Formalin-inactivated vaccine provokes cross-protective immunity in a mouse model of human enterovirus 71 infection","volume":"29","author":"Bek","year":"2011","journal-title":"Vaccine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2406","DOI":"10.1016\/j.vaccine.2014.03.012","article-title":"A combination vaccine comprising of inactivated enterovirus 71 and coxsackievirus A16 elicits balanced protective immunity against both viruses","volume":"32","author":"Cai","year":"2014","journal-title":"Vaccine"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1671","DOI":"10.1016\/j.micinf.2006.01.021","article-title":"Protection of neonatal mice from lethal enterovirus 71 infection by maternal immunization with attenuated Salmonella enterica serovar Typhimurium expressing VP1 of enterovirus 71","volume":"8","author":"Chiu","year":"2006","journal-title":"Microbes Infect."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"9386","DOI":"10.1128\/JVI.02856-06","article-title":"An attenuated strain of enterovirus 71 belonging to genotype a showed a broad spectrum of antigenicity with attenuated neurovirulence in cynomolgus monkeys","volume":"81","author":"Arita","year":"2007","journal-title":"J. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.1016\/j.vaccine.2008.01.058","article-title":"Immunization with virus-like particles of enterovirus 71 elicits potent immune responses and protects mice against lethal challenge","volume":"26","author":"Chung","year":"2008","journal-title":"Vaccine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1016\/S0264-410X(01)00385-1","article-title":"Protection against lethal enterovirus 71 infection in newborn mice by passive immunization with subunit VP1 vaccines and inactivated virus","volume":"20","author":"Wu","year":"2001","journal-title":"Vaccine"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"6951","DOI":"10.1016\/j.vaccine.2010.08.052","article-title":"Enterovirus 71 virus-like particle vaccine: Improved production conditions for enhanced yield","volume":"28","author":"Chung","year":"2010","journal-title":"Vaccine"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"5779","DOI":"10.1016\/j.vaccine.2015.09.043","article-title":"A bivalent virus-like particle based vaccine induces a balanced antibody response against both enterovirus 71 and norovirus in mice","volume":"33","author":"Wang","year":"2015","journal-title":"Vaccine"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1299","DOI":"10.1016\/j.micinf.2007.06.002","article-title":"Passive protection against lethal enterovirus 71 infection in newborn mice by neutralizing antibodies elicited by a synthetic peptide","volume":"9","author":"Foo","year":"2007","journal-title":"Microbes Infect."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"7444","DOI":"10.1016\/j.vaccine.2010.08.080","article-title":"Combined peptides of human enterovirus 71 protect against virus infection in mice","volume":"28","author":"Liu","year":"2010","journal-title":"Vaccine"},{"key":"ref_18","first-page":"712","article-title":"[Mechanism of intravenous immunoglobulin therapy for severe hand-foot-mouth disease: A review]","volume":"27","author":"Cao","year":"2011","journal-title":"Sheng Wu Gong Cheng Xue Bao"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.addr.2016.11.004","article-title":"The state-of-play and future of antibody therapeutics","volume":"122","author":"Elgundi","year":"2017","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Lim, X.F., Jia, Q., Khong, W.X., Yan, B., Premanand, B., Alonso, S., Chow, V.T., and Kwang, J. (2012). Characterization of an isotype-dependent monoclonal antibody against linear neutralizing epitope effective for prophylaxis of enterovirus 71 infection. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0029751"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Kiener, T.K., Jia, Q., Meng, T., Chow, V.T., and Kwang, J. (2014). A novel universal neutralizing monoclonal antibody against enterovirus 71 that targets the highly conserved \u201cknob\u201d region of VP3 protein. PLoS Negl. Trop. Dis., 8.","DOI":"10.1371\/journal.pntd.0002895"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1186\/1743-422X-7-106","article-title":"Monoclonal antibody induced with inactived EV71-Hn2 virus protects mice against lethal EV71-Hn2 virus infection","volume":"7","author":"Chang","year":"2010","journal-title":"Virol. J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4362","DOI":"10.1016\/j.vaccine.2011.04.010","article-title":"Identification and characterization of a cross-neutralization epitope of Enterovirus 71","volume":"29","author":"Liu","year":"2011","journal-title":"Vaccine"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"7663","DOI":"10.1007\/s00253-015-6652-8","article-title":"Generation and characterization of a protective mouse monoclonal antibody against human enterovirus 71","volume":"99","author":"Deng","year":"2015","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1007\/s12250-015-3573-8","article-title":"Monoclonal neutralizing antibodies against EV71 screened from mice immunized with yeast-produced virus-like particles","volume":"30","author":"Lin","year":"2015","journal-title":"Virol. Sin."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"12084","DOI":"10.1128\/JVI.02189-15","article-title":"Single Neutralizing Monoclonal Antibodies Targeting the VP1 GH Loop of Enterovirus 71 Inhibit both Virus Attachment and Internalization during Viral Entry","volume":"89","author":"Ku","year":"2015","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Ye, X., Fan, C., Ku, Z., Zuo, T., Kong, L., Zhang, C., Shi, J., Liu, Q., Chen, T., and Zhang, Y. (2016). Structural Basis for Recognition of Human Enterovirus 71 by a Bivalent Broadly Neutralizing Monoclonal Antibody. PLoS Pathog., 12.","DOI":"10.1371\/journal.ppat.1005454"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Chen, Q., and Davis, K.R. (2016). The potential of plants as a system for the development and production of human biologics. F1000Reserch, 5.","DOI":"10.12688\/f1000research.8010.1"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2419","DOI":"10.1073\/pnas.0914503107","article-title":"Monoclonal antibody produced in plants efficiently treats West Nile virus infection in mice","volume":"107","author":"Lai","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1002\/bit.22652","article-title":"High-level rapid production of full-size monoclonal antibodies in plants by a single-vector DNA replicon system","volume":"106","author":"Huang","year":"2010","journal-title":"Biotechnol. Bioeng."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2055","DOI":"10.1096\/fj.12-219964","article-title":"Production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans","volume":"27","author":"Both","year":"2013","journal-title":"FASEB J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1586\/14760584.2016.1138862","article-title":"EV71 vaccine, a new tool to control outbreaks of hand, foot and mouth disease (HFMD)","volume":"15","author":"Mao","year":"2016","journal-title":"Expert Rev. Vaccines"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1510","DOI":"10.1186\/s40064-016-3074-8","article-title":"A synthetic biology standard for Chinese Hamster Ovary cell genome monitoring and contaminant detection by polymerase chain reaction","volume":"5","author":"Templar","year":"2016","journal-title":"Springerplus"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1002\/biot.201400428","article-title":"Plant-made vaccines against West Nile virus are potent, safe, and economically feasible","volume":"10","author":"Chen","year":"2015","journal-title":"Biotechnol. J."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1456","DOI":"10.1080\/19420862.2016.1227901","article-title":"Techno-economic analysis of a transient plant-based platform for monoclonal antibody production","volume":"8","author":"Nandi","year":"2016","journal-title":"MAbs"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Chen, Q., Lai, H., Hurtado, J., Stahnke, J., Leuzinger, K., and Dent, M. (2013). Agroinfiltration as an Effective and Scalable Strategy of Gene Delivery for Production of Pharmaceutical Proteins. Adv. Tech. Biol. Med., 1.","DOI":"10.4172\/2379-1764.1000103"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1159\/000084805","article-title":"Clinical trial of a plant-derived antibody on recolonization of mutans streptococci","volume":"39","author":"Weintraub","year":"2005","journal-title":"Caries Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1111\/pbi.12416","article-title":"Regulatory approval and a first-in-human phase I clinical trial of a monoclonal antibody produced in transgenic tobacco plants","volume":"13","author":"Ma","year":"2015","journal-title":"Plant Biotechnol. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"648143","DOI":"10.1155\/2015\/648143","article-title":"Clinical Safety and Immunogenicity of Tumor-Targeted, Plant-Made Id-KLH Conjugate Vaccines for Follicular Lymphoma","volume":"2015","author":"Tuse","year":"2015","journal-title":"Biomed. Res. Int."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Li, Y., Yu, J., Qi, X., and Yan, H. (2019). Monoclonal antibody against EV71 3D(pol) inhibits the polymerase activity of RdRp and virus replication. BMC Immunol., 20.","DOI":"10.1186\/s12865-019-0288-x"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1007\/s00705-018-3731-z","article-title":"Identification and production of mouse scFv to specific epitope of enterovirus-71 virion protein-2 (VP2)","volume":"163","author":"Thanongsaksrikul","year":"2018","journal-title":"Arch. Virol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"46402","DOI":"10.1038\/srep46402","article-title":"Effective in vivo therapeutic IgG antibody against VP3 of enterovirus 71 with receptor-competing activity","volume":"7","author":"Jia","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/j.jviromet.2012.06.025","article-title":"Development of murine monoclonal antibodies with potent neutralization effects on enterovirus 71","volume":"186","author":"Ku","year":"2012","journal-title":"J. Virol. Methods"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Ku, Z., Ye, X., Huang, X., Cai, Y., Liu, Q., Li, Y., Su, Z., and Huang, Z. (2013). Neutralizing antibodies induced by recombinant virus-like particles of enterovirus 71 genotype C4 inhibit infection at pre- and post-attachment steps. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0057601"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2335","DOI":"10.1016\/j.vaccine.2015.03.034","article-title":"High-yield production of recombinant virus-like particles of enterovirus 71 in Pichia pastoris and their protective efficacy against oral viral challenge in mice","volume":"33","author":"Zhang","year":"2015","journal-title":"Vaccine"}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/8\/12\/560\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:39:05Z","timestamp":1760189945000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/8\/12\/560"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,12,1]]},"references-count":45,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["plants8120560"],"URL":"https:\/\/doi.org\/10.3390\/plants8120560","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,12,1]]}}}