Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.author | 김승철 | * |
dc.contributor.author | 주웅 | * |
dc.contributor.author | 김윤환 | * |
dc.date.accessioned | 2017-02-15T08:02:03Z | - |
dc.date.available | 2017-02-15T08:02:03Z | - |
dc.date.issued | 2017 | * |
dc.identifier.issn | 1046-5928 | * |
dc.identifier.issn | 1096-0279 | * |
dc.identifier.other | OAK-20080 | * |
dc.identifier.uri | https://dspace.ewha.ac.kr/handle/2015.oak/234510 | - |
dc.description.abstract | Human 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.language | English | * |
dc.publisher | ACADEMIC PRESS INC ELSEVIER SCIENCE | * |
dc.subject | Human papillomavirus | * |
dc.subject | E6 protein | * |
dc.subject | Serology | * |
dc.subject | Cervical cancer | * |
dc.subject | Glutathione S-transferase | * |
dc.title | Two-step chromatographic purification of glutathione S-transferase-tagged human papillomavirus type 16 E6 protein and its application for serology | * |
dc.type | Article | * |
dc.relation.volume | 132 | * |
dc.relation.index | SCIE | * |
dc.relation.index | SCOPUS | * |
dc.relation.startpage | 19 | * |
dc.relation.lastpage | 26 | * |
dc.relation.journaltitle | PROTEIN EXPRESSION AND PURIFICATION | * |
dc.identifier.doi | 10.1016/j.pep.2017.01.004 | * |
dc.identifier.wosid | WOS:000401298600003 | * |
dc.identifier.scopusid | 2-s2.0-85009223601 | * |
dc.author.google | Xu, Mei Ling | * |
dc.author.google | Kim, Seung Cheol | * |
dc.author.google | Kim, Hyoung Jin | * |
dc.author.google | Ju, Woong | * |
dc.author.google | Kim, Yun Hwan | * |
dc.author.google | Kim, Hong-Jin | * |
dc.contributor.scopusid | 김승철(35264000100) | * |
dc.contributor.scopusid | 주웅(8873659700) | * |
dc.contributor.scopusid | 김윤환(55763947200) | * |
dc.date.modifydate | 20240220115825 | * |