{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T04:56:54Z","timestamp":1783400214451,"version":"3.54.6"},"reference-count":23,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2020,8,27]],"date-time":"2020-08-27T00:00:00Z","timestamp":1598486400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008771","name":"Animal and Plant Quarantine Agency","doi-asserted-by":"publisher","award":["B-1543386-2020-21-01"],"award-info":[{"award-number":["B-1543386-2020-21-01"]}],"id":[{"id":"10.13039\/501100008771","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>To differentiate foot-and-mouth disease (FMD)-infected animals from vaccinated livestock, non-structural proteins (NSPs) must be removed during the FMD vaccine manufacturing process. Currently, NSPs cannot be selectively removed from FMD virus (FMDV) culture supernatant. Therefore, polyethylene glycol (PEG) is utilized to partially separate FMDV from NSPs. However, some NSPs remain in the FMD vaccine, which after repeated immunization, may elicit NSP antibodies in some livestock. To address this drawback, chloroform at a concentration of more than 2% (v\/v) was found to remove NSP efficiently without damaging the FMDV particles. Contrary to the PEG-treated vaccine that showed positive NSP antibody responses after the third immunization in goats, the chloroform-treated vaccine did not induce NSP antibodies. In addition to this enhanced vaccine purity, this new method using chloroform could maximize antigen recovery and the vaccine production time could be shortened by two days due to omission of the PEG processing phase. To our knowledge, this is the first report to remove NSPs from FMDV culture supernatant by chemical addition. This novel method could revolutionize the conventional processes of FMD vaccine production.<\/jats:p>","DOI":"10.3390\/vaccines8030483","type":"journal-article","created":{"date-parts":[[2020,8,27]],"date-time":"2020-08-27T08:05:18Z","timestamp":1598515518000},"page":"483","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Efficient Removal of Non-Structural Protein Using Chloroform for Foot-and-Mouth Disease Vaccine Production"],"prefix":"10.3390","volume":"8","author":[{"given":"Sun Young","family":"Park","sequence":"first","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"},{"name":"College of Veterinary Medicine, Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jung-Min","family":"Lee","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0600-1816","authenticated-orcid":false,"given":"Ah-Young","family":"Kim","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sang Hyun","family":"Park","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sim-In","family":"Lee","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyejin","family":"Kim","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"},{"name":"College of Veterinary Medicine, Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jae-Seok","family":"Kim","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jong-Hyeon","family":"Park","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3436-8175","authenticated-orcid":false,"given":"Young-Joon","family":"Ko","sequence":"additional","affiliation":[{"name":"Animal and Plant Quarantine Agency, Gimcheon-si 39660, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Choi-Kyu","family":"Park","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/S0168-1702(02)00257-5","article-title":"Molecular basis of pathogenesis of FMDV","volume":"91","author":"Mason","year":"2003","journal-title":"Virus Res."},{"key":"ref_2","unstructured":"Murphy, F.A., Gibbs, E.P.J., Horzinek, M.C., and Studdert, M.J. (1999). Veterinary Virology, Elsevier."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1128\/CMR.17.2.465-493.2004","article-title":"Foot-and-mouth disease","volume":"17","author":"Grubman","year":"2004","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1080\/07388551.2018.1554619","article-title":"Advances in novel vaccines for foot and mouth disease: Focus on recombinant empty capsids","volume":"39","author":"Mignaqui","year":"2019","journal-title":"Crit. Rev. Biotechnol."},{"key":"ref_5","unstructured":"International Committee, Biological Standards Commission, International Office of Epizootics (2018). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals: Mammals, Birds, and Bees, Renouf Publishing Co. Ltd.. Available online: https:\/\/www.oie.int\/fileadmin\/Home\/eng\/Health_standards\/tahm\/3.01.08_FMD.pdf."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1007\/BF01258364","article-title":"Immunochemical studies of foot-and-mouth disease. VII. Characterization of foot-and-mouth disease virus concentrated by polyethylene glycol precipitation","volume":"30","author":"Wagner","year":"1970","journal-title":"Arch. Fur Die Gesamte Virusforsch."},{"key":"ref_7","first-page":"241","article-title":"Development of an inhibition ELISA test for the detection of non-capsid polyprotein 3ABC in viral suspensions destined for inactivated foot-and-mouth disease vaccines","volume":"126","author":"Bergmann","year":"2006","journal-title":"Dev. Biol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1111\/tbed.12521","article-title":"Global Foot-and-Mouth Disease Research Update and Gap Analysis: 3-Vaccines","volume":"63","author":"Robinson","year":"2016","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.vetmic.2005.12.017","article-title":"Presence of antibodies to non-structural proteins of foot-and-mouth disease virus in repeatedly vaccinated cattle","volume":"115","author":"Lee","year":"2006","journal-title":"Vet. Microbiol."},{"key":"ref_10","unstructured":"Lee, E.G., Park, S.Y., Tark, D.S., Lee, H.S., Ko, Y.J., Kim, Y.G., Park, S.Y., Kim, C.S., Ryu, K.H., and Lee, M.H. (2017). Novel BHK-21 Cell Line Available for Suspension Culture in Serum-Free Medium and Method for Foot-and-Mouth Disease Vaccine Production Using the Same. (KR101812223B1), Available online: https:\/\/patents.google.com\/patent\/KR101812223B1\/en."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1007\/BF01863914","article-title":"Beitrag zur kollektiven behandlung pharmakologischer reihenversuche","volume":"162","author":"Karber","year":"1931","journal-title":"Arch. Fur Exp. Pathol. Phamakologie"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1111\/avj.12190","article-title":"Development and validation of a 3ABC antibody ELISA in Australia for foot and mouth disease","volume":"92","author":"Colling","year":"2014","journal-title":"Aust. Vet. J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1016\/j.virol.2006.02.042","article-title":"Differential distribution of non-structural proteins of foot-and-mouth disease virus in BHK-21 cells","volume":"349","author":"Rosas","year":"2006","journal-title":"Virology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"149","DOI":"10.2147\/OAJCT.S11915","article-title":"Time will tell: Community acceptability of HIV vaccine research before and after the \u201cStep Study\u201d vaccine discontinuation","volume":"2010","author":"Frew","year":"2010","journal-title":"Open Access J. Clin. Trials"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1007\/s00705-016-2805-z","article-title":"The carboxy-terminal half of nonstructural protein 3A is not essential for foot-and-mouth disease virus replication in cultured cell lines","volume":"161","author":"Behura","year":"2016","journal-title":"Arch. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1882","DOI":"10.1016\/j.vaccine.2014.01.067","article-title":"Protection to homologous and heterologous challenge in pigs immunized with vaccine against foot-and-mouth disease type O caused an epidemic in East Asia during 2010\/2011","volume":"32","author":"Park","year":"2014","journal-title":"Vaccine"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"3435","DOI":"10.1016\/j.vaccine.2010.02.072","article-title":"A recombinant truncated FMDV 3AB protein used to better distinguish between infected and vaccinated cattle","volume":"28","author":"He","year":"2010","journal-title":"Vaccine"},{"key":"ref_18","unstructured":"International Committee, Biological Standards Commission, International Office of Epizootics (2012). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals: Mammals, Birds, and Bees, Renouf Publishing Co. Ltd.. Available online: https:\/\/www.oie.int\/doc\/ged\/D12008.pdf."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Feng, X., Ma, J.-W., Sun, S.-Q., Guo, H.-C., Yang, Y.-M., Jin, Y., Zhou, G.-Q., He, J.-J., Guo, J.-H., and Qi, S.-y. (2016). Quantitative detection of the foot-and-mouth disease virus serotype O 146S antigen for vaccine production using a double-antibody sandwich ELISA and nonlinear standard curves. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0149569"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"117","DOI":"10.20506\/rst.26.1.1733","article-title":"Antigen and vaccine banks: Technical requirements and the role of the european antigen bank in emergency foot and mouth disease vaccination","volume":"26","author":"Lombard","year":"2007","journal-title":"Rev. Sci. Tech."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.pep.2015.04.011","article-title":"A hydrophobic interaction chromatography strategy for purification of inactivated foot-and-mouth disease virus","volume":"113","author":"Li","year":"2015","journal-title":"Protein Expr. Purif."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.jchromb.2016.12.033","article-title":"Purification of foot-and-mouth disease virus by heparin as ligand for certain strains","volume":"1049","author":"Du","year":"2017","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.biologicals.2015.06.001","article-title":"Engineering foot-and-mouth disease virus serotype O IND R2\/1975 for one-step purification by immobilized metal affinity chromatography","volume":"43","author":"Biswal","year":"2015","journal-title":"Biologicals"}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/8\/3\/483\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:07:43Z","timestamp":1760177263000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/8\/3\/483"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,27]]},"references-count":23,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["vaccines8030483"],"URL":"https:\/\/doi.org\/10.3390\/vaccines8030483","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,27]]}}}