{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,6]],"date-time":"2026-02-06T23:06:57Z","timestamp":1770419217871,"version":"3.49.0"},"reference-count":62,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2022,10,26]],"date-time":"2022-10-26T00:00:00Z","timestamp":1666742400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Technology","award":["MOST109-2622-B-038-005"],"award-info":[{"award-number":["MOST109-2622-B-038-005"]}]},{"name":"Ministry of Science and Technology","award":["MOST 109-2320-B-075A-001"],"award-info":[{"award-number":["MOST 109-2320-B-075A-001"]}]},{"name":"Ministry of Science and Technology","award":["MOST 109-2320-B-038-053"],"award-info":[{"award-number":["MOST 109-2320-B-038-053"]}]},{"name":"Ministry of Science and Technology","award":["MOST 110-2320-B-075A-001"],"award-info":[{"award-number":["MOST 110-2320-B-075A-001"]}]},{"name":"Ministry of Science and Technology","award":["MOHW110-TDU-B-212-144014"],"award-info":[{"award-number":["MOHW110-TDU-B-212-144014"]}]},{"name":"Ministry of Health and Welfare","award":["MOST109-2622-B-038-005"],"award-info":[{"award-number":["MOST109-2622-B-038-005"]}]},{"name":"Ministry of Health and Welfare","award":["MOST 109-2320-B-075A-001"],"award-info":[{"award-number":["MOST 109-2320-B-075A-001"]}]},{"name":"Ministry of Health and Welfare","award":["MOST 109-2320-B-038-053"],"award-info":[{"award-number":["MOST 109-2320-B-038-053"]}]},{"name":"Ministry of Health and Welfare","award":["MOST 110-2320-B-075A-001"],"award-info":[{"award-number":["MOST 110-2320-B-075A-001"]}]},{"name":"Ministry of Health and Welfare","award":["MOHW110-TDU-B-212-144014"],"award-info":[{"award-number":["MOHW110-TDU-B-212-144014"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Enterovirus 71 (EV71) is the major etiological agent contributing to the development of hand-foot-mouth disease (HFMD). There are not any global available vaccines or antibody drugs against EV71 released yet. In this study, we perform the virus immunization in a cost-effective and convenient approach by preparing virus particles from size exclusion and immunization of chicken. Polyclonal yolk-immunoglobulin (IgY) was simply purified from egg yolk and monoclonal single-chain variable fragments (scFv) were selected via phage display technology with two scFv libraries containing 6.0 \u00d7 106 and 1.3 \u00d7 107 transformants. Specific clones were enriched after 5 rounds of bio-panning and four identical genes were classified after the sequence analysis. Moreover, the higher mutation rates were revealed in the CDR regions, especially in the CDR3. IgY showed specific binding activities to both EV71-infected and Coxsackievirus 16-infected cell lysates and high infectivity inhibitory activity of EV71. However, while IgY detected a 37 kDa protein, the selected scFv seemingly detected higher size proteins which could be cell protein instead of EV71 proteins. Despite the highly effective chicken antibody generation, the purity of virus particles prepared by size exclusion is the limitation of this study, and further characterization should be carried out rigorously.<\/jats:p>","DOI":"10.3390\/ijms232112967","type":"journal-article","created":{"date-parts":[[2022,10,26]],"date-time":"2022-10-26T09:59:37Z","timestamp":1666778377000},"page":"12967","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3412-7400","authenticated-orcid":false,"given":"Chi-Hsin","family":"Lee","sequence":"first","affiliation":[{"name":"Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"},{"name":"School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peng-Nien","family":"Huang","sequence":"additional","affiliation":[{"name":"Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan 333423, Taiwan"},{"name":"Division of Infectious Diseases, Department of Pediatrics, Linkou Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9260-8467","authenticated-orcid":false,"given":"Pharaoh Fellow","family":"Mwale","sequence":"additional","affiliation":[{"name":"Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"},{"name":"School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5452-5537","authenticated-orcid":false,"given":"Wei-Chu","family":"Wang","sequence":"additional","affiliation":[{"name":"Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sy-Jye","family":"Leu","sequence":"additional","affiliation":[{"name":"Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 110301, Taiwan"},{"name":"Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sung-Nien","family":"Tseng","sequence":"additional","affiliation":[{"name":"Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan 333423, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shin-Ru","family":"Shih","sequence":"additional","affiliation":[{"name":"Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan 333423, Taiwan"},{"name":"Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan 333423, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4846-1837","authenticated-orcid":false,"given":"Liao-Chun","family":"Chiang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, National Tsing Hua University, Hsinchu 300040, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8561-0582","authenticated-orcid":false,"given":"Yan-Chiao","family":"Mao","sequence":"additional","affiliation":[{"name":"Division of Clinical Toxicology, Department of Emergency Medicine, Taichung Veterans General Hospital, Taichung 407219, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bor-Yu","family":"Tsai","sequence":"additional","affiliation":[{"name":"Navi Bio-Therapeutics Inc., Taipei 10351, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nhlanhla Benedict","family":"Dlamini","sequence":"additional","affiliation":[{"name":"Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tien-Cuong","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chen-Hsin","family":"Tsai","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Taipei Medical University Hospital, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2534-8436","authenticated-orcid":false,"given":"Yi-Yuan","family":"Yang","sequence":"additional","affiliation":[{"name":"Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"},{"name":"School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan"},{"name":"Core Laboratory of Antibody Generation and Research, Taipei Medical University, Taipei 110301, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,26]]},"reference":[{"key":"ref_1","unstructured":"(2022, March 10). Picornaviridae. Available online: https:\/\/www.picornaviridae.com\/."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/j.forsciint.2016.09.014","article-title":"Coxsackievirus B heart infections and their putative contribution to sudden unexpected death: An 8-year review of patients and victims in the coastal region of Tunisia","volume":"268","author":"Gaaloul","year":"2016","journal-title":"Forensic. Sci. Int."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Royston, L., and Tapparel, C. (2016). Rhinoviruses and Respiratory Enteroviruses: Not as Simple as ABC. Viruses, 8.","DOI":"10.3390\/v8010016"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2645","DOI":"10.1007\/s00705-018-3878-7","article-title":"High frequency of Polio-like Enterovirus C strains with differential clustering of CVA-13 and EV-C99 subgenotypes in a cohort of Malawian children","volume":"163","author":"Brouwer","year":"2018","journal-title":"Arch. Virol."},{"key":"ref_5","unstructured":"King, A., Adams, M.J., Carstens, E.B., and Lefkowitz, E.J. (2011). Virus Taxonomy: Classification and Nomenclature of Viruses, Springer. Ninth Report of The International Committee on Taxonomy of Viruses."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s00705-012-1476-7","article-title":"Molecular identification of enteroviruses associated with aseptic meningitis in children from India","volume":"158","author":"Kumar","year":"2013","journal-title":"Arch. Virol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"850","DOI":"10.1086\/319255","article-title":"Altered cellular but not humoral reactions in children with complicated enterovirus 71 infections in Taiwan","volume":"183","author":"Yang","year":"2001","journal-title":"J. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1097\/01.inf.0000227959.89339.c3","article-title":"Genetic diversity of enterovirus 71 associated with hand, foot and mouth disease epidemics in Japan from 1983 to 2003","volume":"25","author":"Hosoya","year":"2006","journal-title":"Pediatr. Infect. Dis. J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1007\/BF01317504","article-title":"Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria","volume":"60","author":"Chumakov","year":"1979","journal-title":"Arch. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1007\/BF01314873","article-title":"Virological diagnosis of enterovirus type 71 infections: Experiences gained during an epidemic of acute CNS diseases in Hungary in 1978","volume":"71","author":"Nagy","year":"1982","journal-title":"Arch. Virol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/S0168-1702(00)00162-3","article-title":"Genetic analysis of enterovirus 71 isolated from fatal and non-fatal cases of hand, foot and mouth disease during an epidemic in Taiwan, 1998","volume":"68","author":"Shih","year":"2000","journal-title":"Virus Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1056\/NEJM199909233411301","article-title":"An epidemic of enterovirus 71 infection in Taiwan. Taiwan Enterovirus Epidemic Working Group","volume":"341","author":"Ho","year":"1999","journal-title":"N. Engl. J. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1038\/nsmb.2255","article-title":"A sensor-adaptor mechanism for enterovirus uncoating from structures of EV71","volume":"19","author":"Wang","year":"2012","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e1076","DOI":"10.1016\/j.ijid.2010.07.006","article-title":"The largest outbreak of hand; foot and mouth disease in Singapore in 2008: The role of enterovirus 71 and coxsackievirus A strains","volume":"14","author":"Wu","year":"2010","journal-title":"Int. J. Infect. Dis."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Thoa le, P.K., Chiang, P.S., Khanh, T.H., Luo, S.T., Dan, T.N., Wang, Y.F., Thuong, T.C., Chung, W.Y., Hung, N.T., and Wang, J.R. (2013). Genetic and antigenic characterization of enterovirus 71 in Ho Chi Minh City, Vietnam, 2011. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069895"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"868","DOI":"10.1016\/S1474-4422(08)70207-2","article-title":"Enterovirus 71 infection: A new threat to global public health?","volume":"7","author":"Qiu","year":"2008","journal-title":"Lancet Neurol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1056\/NEJMoa1304923","article-title":"Efficacy, safety, and immunogenicity of an enterovirus 71 vaccine in China","volume":"370","author":"Zhu","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"395","DOI":"10.3345\/kjp.2016.59.10.395","article-title":"Enterovirus 71 infection and neurological complications","volume":"59","author":"Lee","year":"2016","journal-title":"Korean J. Pediatr."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Liu, C.-C., Guo, M.-S., Lin, F.H.-Y., Hsiao, K.-N., Chang, K.H.-W., Chou, A.-H., Wang, Y.-C., Chen, Y.-C., Yang, C.-S., and Chong, P.C.-S. (2011). Purification and characterization of enterovirus 71 viral particles produced from vero cells grown in a serum-free microcarrier bioreactor system. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0020005"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1007\/s12250-018-0071-9","article-title":"Recent progress on functional genomics research of enterovirus 71","volume":"34","author":"Wang","year":"2019","journal-title":"Virol. Sin."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"9743","DOI":"10.1128\/jvi.71.12.9743-9752.1997","article-title":"Dissecting the roles of VP0 cleavage and RNA packaging in picornavirus capsid stabilization: The structure of empty capsids of foot-and-mouth disease virus","volume":"71","author":"Curry","year":"1997","journal-title":"J. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1038\/nri2744","article-title":"Monoclonal antibodies: Versatile platforms for cancer immunotherapy","volume":"10","author":"Weiner","year":"2010","journal-title":"Nat. Rev. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.vetimm.2009.12.011","article-title":"IgY: A promising antibody for use in immunodiagnostic and in immunotherapy","volume":"135","author":"Tambourgi","year":"2010","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_24","first-page":"75","article-title":"Avian egg yolk antibodies. The egg laying capacity of hens following immunisation with antigens of different kind and origin and the efficiency of egg yolk antibodies in comparison to mammalian antibodies","volume":"11","author":"Schade","year":"1994","journal-title":"Altex"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1146\/annurev-food-022811-101137","article-title":"Egg yolk antibodies for passive immunity","volume":"3","author":"Mine","year":"2012","journal-title":"Annu. Rev. Food Sci. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/S0022-1759(00)00221-0","article-title":"Methods for the generation of chicken monoclonal antibody fragments by phage display","volume":"242","author":"Rader","year":"2000","journal-title":"J. Immunol. Methods"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12929-016-0223-x","article-title":"Advancement and applications of peptide phage display technology in biomedical science","volume":"23","author":"Wu","year":"2016","journal-title":"J. Biomed. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"8875","DOI":"10.1007\/s00253-019-10145-1","article-title":"A microbial expression system for high-level production of scFv HIV-neutralizing antibody fragments in Escherichia coli","volume":"103","author":"Petrus","year":"2019","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Lee, C.-H., Lee, Y.-C., Lee, Y.-L., Leu, S.-J., Lin, L.-T., Chen, C.-C., Chiang, J.-R., Mwale, P.F., Tsai, B.-Y., and Hung, C.-S. (2017). Single chain antibody fragment against venom from the snake daboia russelii formosensis. Toxins, 9.","DOI":"10.3390\/toxins9110347"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Leu, S.J., Lee, Y.C., Lee, C.H., Liao, P.Y., Chiang, C.W., Yang, C.M., Su, C.H., Ou, T.Y., Liu, K.J., and Lo, H.J. (2020). Generation and Characterization of Single Chain Variable Fragment against Alpha-Enolase of Candida albicans. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21082903"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Mwale, P.F., Lee, C.H., Huang, P.N., Tseng, S.N., Shih, S.R., Huang, H.Y., Leu, S.J., Huang, Y.J., Chiang, L.C., and Mao, Y.C. (2021). In Vitro Characterization of Neutralizing Hen Antibodies to Coxsackievirus A16. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22084146"},{"key":"ref_32","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_33","doi-asserted-by":"crossref","unstructured":"Brouwer, L., Moreni, G., Wolthers, K.C., and Pajkrt, D. (2021). World-Wide Prevalence and Genotype Distribution of Enteroviruses. Viruses, 13.","DOI":"10.3390\/v13030434"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1016\/j.ajic.2013.01.026","article-title":"Viral outbreaks in neonatal intensive care units: What we do not know","volume":"41","author":"Civardi","year":"2013","journal-title":"Am. J. Infect. Control"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"696003","DOI":"10.3389\/fimmu.2021.696003","article-title":"Immunoglobulin Y for Potential Diagnostic and Therapeutic Applications in Infectious Diseases","volume":"12","author":"Lee","year":"2021","journal-title":"Front Immunol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2528","DOI":"10.1096\/fasebj.4.8.1970792","article-title":"Efficient production of chicken egg yolk antibodies against a conserved mammalian protein","volume":"4","author":"Gassmann","year":"1990","journal-title":"FASEB J."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"892","DOI":"10.1002\/ppul.20875","article-title":"Good effect of IgY against Pseudomonas aeruginosa infections in cystic fibrosis patients","volume":"43","author":"Nilsson","year":"2008","journal-title":"Pediatr. Pulmonol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1275","DOI":"10.1126\/science.2531466","article-title":"Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda","volume":"246","author":"Huse","year":"1989","journal-title":"Science"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"12612","DOI":"10.1073\/pnas.192467999","article-title":"A method for the generation of combinatorial antibody libraries using pIX phage display","volume":"99","author":"Gao","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1146\/annurev.bioeng.2.1.339","article-title":"Antibody engineering","volume":"2","author":"Maynard","year":"2000","journal-title":"Annu. Rev. Biomed. Eng."},{"key":"ref_41","first-page":"17","article-title":"Hybridoma technology for production of monoclonal antibodies","volume":"1","author":"Pandey","year":"2010","journal-title":"Hybridoma"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e17","DOI":"10.1038\/jid.2013.521","article-title":"Antibody phage display: Technique and applications","volume":"134","author":"Hammers","year":"2014","journal-title":"J. Investig. Dermatol."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Vashist, S.K., and Luong, J.H. (2018). Handbook of Immunoassay Technologies: Approaches, Performances, and Applications, Academic Press.","DOI":"10.1016\/B978-0-12-811762-0.05001-5"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1128\/AEM.02608-15","article-title":"Antibodies against Venom of the Snake Deinagkistrodon acutus","volume":"82","author":"Lee","year":"2016","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"3072","DOI":"10.1073\/pnas.87.8.3072","article-title":"Somatic diversification of chicken immunoglobulin light chains by point mutations","volume":"87","author":"Parvari","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/0092-8674(89)90879-9","article-title":"Somatic hyperconversion diversifies the single Vh gene of the chicken with a high incidence in the D region","volume":"59","author":"Reynaud","year":"1989","journal-title":"Cell"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/S1074-7613(00)00006-6","article-title":"Diversity in the CDR3 region of V(H) is sufficient for most antibody specificities","volume":"13","author":"Xu","year":"2000","journal-title":"Immunity"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-018-30322-y","article-title":"Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples","volume":"8","author":"Isaacs","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Saarinen, N.V., Stone, V.M., Hankaniemi, M.M., Mazur, M.A., Vuorinen, T., Flodstr\u00f6m-Tullberg, M., Hy\u00f6ty, H., Hyt\u00f6nen, V.P., and Laitinen, O.H. (2020). Antibody Responses against Enterovirus Proteases are Potential Markers for an Acute Infection. Viruses, 12.","DOI":"10.3390\/v12010078"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Wu, C.Y., Yu, S.L., Chen, Y.T., Chen, Y.H., Hsiao, P.W., Chow, Y.H., and Chen, J.R. (2019). The mature EV71 virion induced a broadly cross-neutralizing VP1 antibody against subtypes of the EV71 virus. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0210553"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"562768","DOI":"10.3389\/fmicb.2020.562768","article-title":"Human Antibodies to VP4 Inhibit Replication of Enteroviruses Across Subgenotypes and Serotypes, and Enhance Host Innate Immunity","volume":"11","author":"Phanthong","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1007\/s40995-020-01025-9","article-title":"Recombinant Human SCARB2 Expressed in Escherichia coli and its Potential in Enterovirus 71 Blockage","volume":"45","author":"Nguyen","year":"2021","journal-title":"Iran. J. Sci. Technol. Trans. A Sci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1186\/s12929-020-0615-9","article-title":"Cellular receptors for enterovirus A71","volume":"27","author":"Kobayashi","year":"2020","journal-title":"J. Biomed. Sci."},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Li, M.L., Shih, S.R., Tolbert, B.S., and Brewer, G. (2021). Enterovirus A71 Vaccines. Vaccines, 9.","DOI":"10.3390\/vaccines9030199"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1186\/s12929-019-0560-7","article-title":"Antivirals and vaccines for Enterovirus A71","volume":"26","author":"Lin","year":"2019","journal-title":"J. Biomed. Sci."},{"key":"ref_56","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_57","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.seppur.2013.09.026","article-title":"An evaluation of virus removal mechanisms by ultrafiltration membranes using MS2 and \u03c6X174 bacteriophage","volume":"120","author":"ElHadidy","year":"2013","journal-title":"Sep. Purif. Technol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.memsci.2011.12.050","article-title":"Mechanisms of virus removal from secondary wastewater effluent by low pressure membrane filtration","volume":"409","author":"Huang","year":"2012","journal-title":"J. Membr. Sci."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/0022-1759(93)90380-P","article-title":"Production and purification of Fab\u2032 fragments from chicken egg yolk immunoglobulin Y (IgY)","volume":"162","author":"Akita","year":"1993","journal-title":"J. Immunol. Methods"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/0022-1759(93)90179-B","article-title":"Comparison of four purification methods for the production of immunoglobulins from eggs laid by hens immunized with an enterotoxigenic E. coli strain","volume":"160","author":"Akita","year":"1993","journal-title":"J. Immunol. Methods"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1556\/EuJMI.4.2014.2.1","article-title":"Production of recombinant antibodies using bacteriophages","volume":"4","author":"Shukra","year":"2014","journal-title":"Eur. J. Microbiol. Immunol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"e01922","DOI":"10.1128\/AAC.01922-19","article-title":"Antiviral Activity of a Llama-Derived Single-Domain Antibody against Enterovirus A71","volume":"64","author":"Huang","year":"2020","journal-title":"Antimicrob. 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