{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T15:00:25Z","timestamp":1787065225229,"version":"build-2736575974"},"reference-count":53,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2015,6,30]],"date-time":"2015-06-30T00:00:00Z","timestamp":1435622400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["MOST 103-2622-E-007-024-CC1"],"award-info":[{"award-number":["MOST 103-2622-E-007-024-CC1"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["NSC 102-2622-E-007-022-CC1"],"award-info":[{"award-number":["NSC 102-2622-E-007-022-CC1"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["NSC 101-2628- E-007-009-MY3"],"award-info":[{"award-number":["NSC 101-2628- E-007-009-MY3"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["NSC 100-2622-E-007-003-CC2"],"award-info":[{"award-number":["NSC 100-2622-E-007-003-CC2"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100008903","name":"Ministry of Health and Welfare, Taiwan","doi-asserted-by":"crossref","award":["DOH98-DC-1013"],"award-info":[{"award-number":["DOH98-DC-1013"]}],"id":[{"id":"10.13039\/100008903","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100008903","name":"Ministry of Health and Welfare, Taiwan","doi-asserted-by":"crossref","award":["DOH99-TD-I-111-TM022"],"award-info":[{"award-number":["DOH99-TD-I-111-TM022"]}],"id":[{"id":"10.13039\/100008903","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100008903","name":"Ministry of Health and Welfare, Taiwan","doi-asserted-by":"crossref","award":["MOHW103-TDU-PB-211-122021"],"award-info":[{"award-number":["MOHW103-TDU-PB-211-122021"]}],"id":[{"id":"10.13039\/100008903","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100008903","name":"Ministry of Health and Welfare, Taiwan","doi-asserted-by":"crossref","award":["MOHW102-TDU-PB-111-NSC108"],"award-info":[{"award-number":["MOHW102-TDU-PB-111-NSC108"]}],"id":[{"id":"10.13039\/100008903","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech &amp; Bioengineering"],"published-print":{"date-parts":[[2015,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:label\/>\n                    <jats:p>\n                      Enterovirus 71 (EV71) is responsible for the outbreaks of hand\u2010foot\u2010and\u2010mouth disease in the Asia\u2010Pacific region. To produce the virus\u2010like particle (VLP) vaccine, we previously constructed recombinant baculoviruses to co\u2010express EV71 P1 polypeptide and 3CD protease using the Bac\u2010to\u2010Bac\n                      <jats:sup>\u00ae<\/jats:sup>\n                      vector system. The recombinant baculoviruses resulted in P1 cleavage by 3CD and subsequent VLP assembly in infected insect cells, but caused either low VLP yield or excessive VLP degradation. To tackle the problems, here we explored various expression cassette designs and flashBAC GOLD\u2122 vector system which was deficient in\n                      <jats:italic>v\u2010cath<\/jats:italic>\n                      and\n                      <jats:italic>chiA<\/jats:italic>\n                      genes. We found that the recombinant baculovirus constructed using the flashBAC GOLD\u2122 system was insufficient to improve the EV71 VLP yield. Nonetheless, BacF\u2010P1\u2010C3CD, a recombinant baculovirus constructed using the flashBAC GOLD\n                      <jats:sup>TM<\/jats:sup>\n                      system to express P1 under the\n                      <jats:italic>polh<\/jats:italic>\n                      promoter and 3CD under the\n                      <jats:italic>CMV<\/jats:italic>\n                      promoter, dramatically improved the VLP yield while alleviating the VLP degradation. Infection of High Five\n                      <jats:sup>TM<\/jats:sup>\n                      cells with BacF\u2010P1\u2010C3CD enhanced the total and extracellular VLP yield to \u2248268 and \u2248171\u2009mg\/L, respectively, which enabled the release of abundant VLP into the supernatant and simplified the downstream purification. Intramuscular immunization of mice with 5\u2009\u03bcg purified VLP induced cross\u2010protective humoral responses and conferred protection against lethal virus challenge. Given the significantly improved extracellular VLP yield (\u2248171\u2009mg\/L) and the potent immunogenicity conferred by 5\u2009\u03bcg VLP, one liter High Five\n                      <jats:sup>TM<\/jats:sup>\n                      culture produced \u224812,000 doses of purified vaccine, thus rendering the EV71 VLP vaccine economically viable and able to compete with inactivated virus vaccines. Biotechnol. Bioeng. 2015;112: 2005\u20132015. \u00a9 2015 Wiley Periodicals, Inc.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/bit.25625","type":"journal-article","created":{"date-parts":[[2015,5,21]],"date-time":"2015-05-21T19:33:05Z","timestamp":1432236785000},"page":"2005-2015","source":"Crossref","is-referenced-by-count":16,"title":["Enhanced enterovirus 71 virus\u2010like particle yield from a new baculovirus design"],"prefix":"10.1002","volume":"112","author":[{"given":"Shih\u2010Yeh","family":"Lin","sequence":"first","affiliation":[{"name":"Department of Chemical Engineering National Tsing Hua University Hsinchu Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chia\u2010Tsui","family":"Yeh","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine National Defense Medical Center Taipei Taiwan"},{"name":"Department of Life Science National Taiwan Normal University Taipei Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wan\u2010Hua","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering National Tsing Hua University Hsinchu Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng\u2010Ping","family":"Yu","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine National Defense Medical Center Taipei Taiwan"},{"name":"Department of Pathology Tri\u2010Service General Hospital National Defense Medical Center Taipei Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen\u2010Chin","family":"Lin","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine National Defense Medical Center Taipei Taiwan"},{"name":"Department of Veterinary Medicine College of Veterinary Medicine National Chung Hsing University Taichung Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jyh\u2010Yuan","family":"Yang","sequence":"additional","affiliation":[{"name":"Center for Research Diagnostics and Vaccine Development Centers for Disease Control Taipei Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hsueh\u2010Ling","family":"Wu","sequence":"additional","affiliation":[{"name":"Institute of Preventive Medicine National Defense Medical Center Taipei Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu\u2010Chen","family":"Hu","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering National Tsing Hua University Hsinchu Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,6,30]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2010.08.005"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1036"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2013.03.015"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2010.08.052"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2008.01.058"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v12.i6.921"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0290(19991020)65:2<133::AID-BIT2>3.0.CO;2-X"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1586\/erv.12.153"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1389-1723(01)80258-9"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1997.8816"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2011.06.011"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10565-009-9133-y"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1042\/BA20090130"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2000.0586"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/bp990094k"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1024071514438"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1586\/14760584.7.3.363"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2004.07.006"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1586\/erv.13.18"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.200900267"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12033-010-9268-3"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1517\/14712598.2014.935330"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2013.05.019"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jit429"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/cti.2014.24"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2015.04.077"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiosc.2013.08.015"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1586\/14760584.2014.951637"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0111496"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2011.12.081"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0020005"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2010.08.080"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.25159"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M1114.624536"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2007.05.022"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-002-0976-x"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2004.09.009"},{"key":"e_1_2_8_39_1","doi-asserted-by":"crossref","first-page":"6619","DOI":"10.1128\/jvi.68.10.6619-6625.1994","article-title":"A cysteine protease encoded by the baculovirus Bombyx mori nuclear polyhedrosis virus","volume":"68","author":"Ohkawa T","year":"1994","journal-title":"J Virol"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1218713"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1042\/bst0270928"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2009.04.004"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2014.130"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M309351200"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-69-4-765"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0094401"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2009.06.012"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/nsmb.2255"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.78.15.7916-7924.2004"},{"key":"e_1_2_8_50_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2006.03.021"},{"key":"e_1_2_8_51_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2013.11.069"},{"key":"e_1_2_8_52_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2180-13-287"},{"key":"e_1_2_8_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(12)61764-4"},{"key":"e_1_2_8_54_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(13)61049-1"}],"container-title":["Biotechnology and Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fbit.25625","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bit.25625","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T17:55:59Z","timestamp":1759946159000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/bit.25625"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,6,30]]},"references-count":53,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2015,10]]}},"alternative-id":["10.1002\/bit.25625"],"URL":"https:\/\/doi.org\/10.1002\/bit.25625","archive":["Portico"],"relation":{},"ISSN":["0006-3592","1097-0290"],"issn-type":[{"value":"0006-3592","type":"print"},{"value":"1097-0290","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,6,30]]}}}