{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,4,9]],"date-time":"2025-04-09T06:08:52Z","timestamp":1744178932055,"version":"3.37.3"},"reference-count":29,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2015,7,6]],"date-time":"2015-07-06T00:00:00Z","timestamp":1436140800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"National \u201c863\u201d High-Tech Program of China","award":["SS2012AA020905"],"award-info":[{"award-number":["SS2012AA020905"]}]},{"name":"China Key Project of New Medicine Development","award":["2014ZX09304314-003"],"award-info":[{"award-number":["2014ZX09304314-003"]}]},{"DOI":"10.13039\/501100001809","name":"China National Natural Science Foundation","doi-asserted-by":"crossref","award":["81373109"],"award-info":[{"award-number":["81373109"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"China Jiangsu Province Natural Science Foundation","award":["BK2012526"],"award-info":[{"award-number":["BK2012526"]}]},{"name":"China Jiangsu Province Social Development Foundation","award":["BE2010603"],"award-info":[{"award-number":["BE2010603"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotech and App Biochem"],"published-print":{"date-parts":[[2016,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Glycoprotein D (gD2) is the most important candidate antigen for herpes simplex virus type 2 (HSV\u20102) vaccine development. Establishment of a stable eukaryotic cell line to overexpress gD2 and an efficient purification process to purify is essential for the development of subunit vaccine against HSV\u20102. The DNA sequence of the extracellular epitope\u2010rich fragment of gD2 was optimized, chemically synthesized, and cloned into plasmid pMD902. The recombinant plasmid pMD902\u2010gD was stably transfected into CHO\u2010DG44 cells, and cell lines with high levels of expression of gD2 were established. The recombinant gD2 was purified efficiently using an anion exchange column and a Sephadex G\u201025 desalting column. The yield of the purified gD2 was 57\u00a0mg\/L of serum\u2010free culture medium, and its purity was determined to be about 95% by HPLC analysis. Finally, the immunogenicity of the purified gD2 was measured and it induced strong and specific humoral immunity and higher level of cellular immune response than gD2 expressed in prokaryotic cells. We established a stable, secretory, and high\u2010yield gD2\u2010expression cell line and an easy and efficient gD2\u2010purification process, which lays the foundation for preparation of large amount of gD2 that is essential for HSV\u20102 subunit vaccine development.<\/jats:p>","DOI":"10.1002\/bab.1386","type":"journal-article","created":{"date-parts":[[2015,4,23]],"date-time":"2015-04-23T10:31:44Z","timestamp":1429785104000},"page":"312-319","source":"Crossref","is-referenced-by-count":1,"title":["Overexpression and purification of HSV\u20102 glycoprotein D in suspension CHO cells with serum\u2010free medium and immunogenicity analysis"],"prefix":"10.1002","volume":"63","author":[{"given":"Yong","family":"Qi","sequence":"first","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques  Nanjing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yueyue","family":"Xu","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques  Nanjing People's Republic of China"},{"name":"Department of Biochemistry and Molecular Biology School of Preclinical Medicine Nanjing Medical University  Nanjing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ying","family":"Pan","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques  Nanjing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Suqin","family":"Li","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques  Nanjing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bingjun","family":"Li","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques  Nanjing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mingjie","family":"Pan","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques  Nanjing People's Republic of China"},{"name":"Department of Biochemistry and Molecular Biology School of Preclinical Medicine Nanjing Medical University  Nanjing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shumin","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Institutes for Food and Drug Control  Beijing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuexi","family":"Li","sequence":"additional","affiliation":[{"name":"Huadong Research Institute for Medicine and Biotechniques  Nanjing People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2015,7,6]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(08)60920-4"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/45.suppl_4.1"},{"key":"e_1_2_7_4_1","doi-asserted-by":"crossref","first-page":"2760","DOI":"10.1128\/JVI.72.4.2760-2764.1998","volume":"72","author":"Lekstrom\u2010Himes J. A.","year":"1998","journal-title":"J. Virol."},{"key":"e_1_2_7_5_1","doi-asserted-by":"crossref","first-page":"9166","DOI":"10.1128\/JVI.72.11.9166-9172.1998","volume":"72","author":"Maggs D. J.","year":"1998","journal-title":"J. Virol."},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1097\/00007435-198510000-00009"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-67-8-1601"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI57148"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2013.08.066"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.63.1.52-58.1989"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.virol.2005.09.016"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/309633a0"},{"key":"e_1_2_7_13_1","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1128\/jvi.62.4.1347-1354.1988","volume":"62","author":"Johnson D. C.","year":"1988","journal-title":"J. Virol."},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/157.5.897"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1086\/429240"},{"key":"e_1_2_7_16_1","first-page":"870","volume":"48","author":"Werner R. G.","year":"1998","journal-title":"Arzneimittel\u2010Forschung"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2005.12.111"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2013.09.064"},{"key":"e_1_2_7_19_1","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1128\/jcm.33.2.381-384.1995","volume":"33","author":"B\u00f6lin I.","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.10449"},{"key":"e_1_2_7_21_1","doi-asserted-by":"crossref","first-page":"3356","DOI":"10.1128\/jvi.61.11.3356-3364.1987","volume":"61","author":"Highlander S. L.","year":"1987","journal-title":"J. Virol."},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.62.12.4605-4612.1988"},{"key":"e_1_2_7_23_1","doi-asserted-by":"crossref","first-page":"2569","DOI":"10.1128\/jvi.64.6.2569-2576.1990","volume":"64","author":"Johnson D. C.","year":"1990","journal-title":"J. Virol."},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1001\/jama.282.4.331"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa011915"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00849-11"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00849-11"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.03163-13"},{"key":"e_1_2_7_29_1","first-page":"59","volume":"12","author":"Fotouhi F.","year":"2008","journal-title":"Iran. Biomed. J."},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0264-410X(98)00434-4"}],"container-title":["Biotechnology and Applied Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fbab.1386","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/bab.1386","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,6]],"date-time":"2023-10-06T14:49:42Z","timestamp":1696603782000},"score":1,"resource":{"primary":{"URL":"https:\/\/iubmb.onlinelibrary.wiley.com\/doi\/10.1002\/bab.1386"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,7,6]]},"references-count":29,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2016,5]]}},"alternative-id":["10.1002\/bab.1386"],"URL":"https:\/\/doi.org\/10.1002\/bab.1386","archive":["Portico"],"relation":{},"ISSN":["0885-4513","1470-8744"],"issn-type":[{"type":"print","value":"0885-4513"},{"type":"electronic","value":"1470-8744"}],"subject":[],"published":{"date-parts":[[2015,7,6]]}}}