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dc.contributor.author김승철-
dc.contributor.author주웅-
dc.contributor.author김윤환-
dc.date.accessioned2017-02-15T08:02:03Z-
dc.date.available2017-02-15T08:02:03Z-
dc.date.issued2017-
dc.identifier.issn1046-5928-
dc.identifier.issn1096-0279-
dc.identifier.otherOAK-20080-
dc.identifier.urihttps://dspace.ewha.ac.kr/handle/2015.oak/234510-
dc.description.abstractHuman papillomavirus (HPV) E6 protein is an oncoprotein with a pivotal role in cervical carcinogenesis. Expression and purification of HPV E6 from Escherichia coli (E. coli) has been difficult because of its strong hydrophobicity even when expressed as a fusion protein with glutathione S-transferase (GST). There has been no protocol suggested for purifying GST-tagged HPV E6 protein with high purity so far. Herein, we provide efficient protocol for purifying GST-HPV16 E6 protein for the first time. In the current study, the GST-tagged protein was expressed in E. coli and a purification method was designed using cation exchange chromatography followed by GST-affinity chromatography. Using physiological pH buffer during cell lysis and first cation-exchange chromatography significantly reduced yield of full-length GST-HPV16 E6 protein. It was found that using an alkaline buffer during cation-exchange chromatography was needed to obtain full length GST-HPV16 E6 protein. GST-HPV16 E6 protein recovered from the purification using alkaline condition retained its inherent p53-binding ability. Moreover, we were able to detect anti-HPV16 E6 antibodies with high sensitivity in sera from patients with cervical cancer using the GST HPV16 E6 protein. It was found that the GST-HPV16 E6 protein could be used as a coating agent to enhance the sensitivity of detection of serum anti-HPV16 E6 antibodies when treated with ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA). These results indicate that the two-step chromatographic purification allows obtaining high purity of GST-HPV16 E6 protein and the GST-HPV16 E6 is suitable to be used as an antigen of serology assay. (C) 2017 Elsevier Inc. All rights reserved.-
dc.languageEnglish-
dc.publisherACADEMIC PRESS INC ELSEVIER SCIENCE-
dc.subjectHuman papillomavirus-
dc.subjectE6 protein-
dc.subjectSerology-
dc.subjectCervical cancer-
dc.subjectGlutathione S-transferase-
dc.titleTwo-step chromatographic purification of glutathione S-transferase-tagged human papillomavirus type 16 E6 protein and its application for serology-
dc.typeArticle-
dc.relation.volume132-
dc.relation.indexSCIE-
dc.relation.indexSCOPUS-
dc.relation.startpage19-
dc.relation.lastpage26-
dc.relation.journaltitlePROTEIN EXPRESSION AND PURIFICATION-
dc.identifier.doi10.1016/j.pep.2017.01.004-
dc.identifier.wosidWOS:000401298600003-
dc.identifier.scopusid2-s2.0-85009223601-
dc.author.googleXu, Mei Ling-
dc.author.googleKim, Seung Cheol-
dc.author.googleKim, Hyoung Jin-
dc.author.googleJu, Woong-
dc.author.googleKim, Yun Hwan-
dc.author.googleKim, Hong-Jin-
dc.contributor.scopusid김승철(35264000100)-
dc.contributor.scopusid주웅(8873659700)-
dc.contributor.scopusid김윤환(55763947200)-
dc.date.modifydate20211117115650-
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의과대학 > 의학과 > Journal papers
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