{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T12:50:36Z","timestamp":1740142236348,"version":"3.37.3"},"reference-count":0,"publisher":"Bentham Science Publishers Ltd.","issue":"11","funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81860257"],"award-info":[{"award-number":["81860257"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["eurekaselect.com"],"crossmark-restriction":true},"short-container-title":["PPL"],"published-print":{"date-parts":[[2020,11,16]]},"abstract":"<jats:sec>\n<jats:title>Background:<\/jats:title>\n<jats:p> Tetanus is an infectious disease caused by Clostridium secreting\ntetanus toxin in anaerobic environment. The fragment C of Tetanus toxin (TTc) has been widely\nstudied as a candidate vaccine to replace the existing tetanus toxoid vaccine.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Objective:<\/jats:title>\n<jats:p>In this study, we established a simple method to purify recombinant protein TTc with\nion-exchange chromatography from Escherichia coli expression systems.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Methods:<\/jats:title>\n<jats:p>The TTc gene sequence was cloned into pET26b (+) vector and transferred to E. coli\nBL21 (DE3) for expression. The fermentation conditions (IPTG concentration, Induction temperature,\nInduction time) were optimized to obtain more soluble proteins. The soluble proteins were\npurified by Anion exchange chromatography and Cation exchange chromatography. The sequence\nof columns in the purification process was discussed. Finally, the stability of purified TTc protein\nwere determined, the secondary structure of the purified TTc protein was determined by circular\ndichroism. The molecular weight of the purified TTc protein was determined by liquid chromatograph-\nmass spectrometer. Furthermore, we verified the immunogenicity of the purified protein in\nmice.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Results:<\/jats:title>\n<jats:p> The purity of TTc improved from 34% to 88% after the first anion exchange column, and\nthe final yield of recombinant TTc (purity &gt; 95%) can reach 84.79% after the following cation\nexchange chromatography. The recombinant TTc had a molecular weight of 51.737 KDa, was\nstable at 4\u00b0C and weak alkaline environment, was a \u03b2-sheet secondary structure, and had strong\nimmunogenicity.<\/jats:p>\n<\/jats:sec>\n<jats:sec>\n<jats:title>Conclusion:<\/jats:title>\n<jats:p>The purification method we developed might be an efficient method for the industrial\nproduction of tetanus recombinant TTc vaccine.<\/jats:p>\n<\/jats:sec>","DOI":"10.2174\/0929866527666200528113327","type":"journal-article","created":{"date-parts":[[2020,5,28]],"date-time":"2020-05-28T15:14:46Z","timestamp":1590678886000},"page":"1132-1140","update-policy":"https:\/\/doi.org\/10.2174\/bsp_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Expression and Purification of Tetanus Toxin Fragment C in Escherichia coli BL21(DE3)"],"prefix":"10.2174","volume":"27","author":[{"given":"Pengdi","family":"Chai","sequence":"first","affiliation":[{"name":"School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiuying","family":"Pu","sequence":"additional","affiliation":[{"name":"School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jianqiang","family":"Li","sequence":"additional","affiliation":[{"name":"Jiangsu Theravac Bio-Pharmaceutical Co., Ltd, Nanjing 210042, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoyu","family":"Xia","sequence":"additional","affiliation":[{"name":"School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Ge","sequence":"additional","affiliation":[{"name":"Jiangsu Theravac Bio-Pharmaceutical Co., Ltd, Nanjing 210042, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amiao","family":"Luo","sequence":"additional","affiliation":[{"name":"School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hui","family":"Su","sequence":"additional","affiliation":[{"name":"School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weijie","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jianzhong","family":"Ma","sequence":"additional","affiliation":[{"name":"School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou 730050, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"965","container-title":["Protein &amp; Peptide Letters"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/eurekaselect.com\/article\/download\/182316","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.eurekaselect.com\/182316\/article","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/eurekaselect.com\/article\/download\/182316","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,5]],"date-time":"2021-07-05T14:00:07Z","timestamp":1625493607000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.eurekaselect.com\/182316\/article"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,16]]},"references-count":0,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2020,11,16]]}},"alternative-id":["LiveAll1"],"URL":"https:\/\/doi.org\/10.2174\/0929866527666200528113327","relation":{},"ISSN":["0929-8665"],"issn-type":[{"type":"print","value":"0929-8665"}],"subject":[],"published":{"date-parts":[[2020,11,16]]},"assertion":[{"value":"Peer Reviewed","order":0,"name":"review_status","label":"Review Status","group":{"name":"peer_review_details","label":"Peer Review Details"}},{"value":"Single blind","order":1,"name":"review_process","label":"Review Process","group":{"name":"peer_review_details","label":"Peer Review Details"}},{"value":"Checked with iThenticate","order":0,"name":"screening_status","label":"Screening Status","group":{"name":"plagiarism_screening","label":"Plagiarism Screening"}},{"value":"2020-01-18","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-04-23","order":1,"name":"revised","label":"Revised","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-04-29","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-11-16","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}