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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.modifydate20240220115825*
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의과대학 > 의학과 > Journal papers
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