DC Field | Value | Language |
---|---|---|
dc.contributor.author | Seo, Hyo-Deok | ko |
dc.contributor.author | Lee, Joong-Jae | ko |
dc.contributor.author | Kim, Yu Jung | ko |
dc.contributor.author | Hantschel, Oliver | ko |
dc.contributor.author | Lee, Seung-Goo | ko |
dc.contributor.author | Kim, Hak-Sung | ko |
dc.date.accessioned | 2017-02-02T02:26:05Z | - |
dc.date.available | 2017-02-02T02:26:05Z | - |
dc.date.created | 2017-01-23 | - |
dc.date.created | 2017-01-23 | - |
dc.date.issued | 2017-01 | - |
dc.identifier.citation | ANALYTICA CHIMICA ACTA, v.950, pp.184 - 191 | - |
dc.identifier.issn | 0003-2670 | - |
dc.identifier.uri | http://hdl.handle.net/10203/220384 | - |
dc.description.abstract | Enzyme-linked immunoassays based on an antibody-antigen interaction are widely used in biological and medical sciences. However, the conjugation of an enzyme to antibodies needs an additional chemical process, usually resulting in randomly cross-linked molecules and a loss of the binding affinity and enzyme activity. Herein, we present the development of an alkaline phosphatase-fused repebody as a new format of immuno-reagent for immunoassays. A repebody specifically binding to human TNF-alpha (hTNF-alpha) was selected through a phage display, and its binding affinity was increased up to 49 nM using a modular engineering approach. A monomeric alkaline phosphatase (mAP), which was previously isolated from a metagenome library, was genetically fused to the repebody as a signal generator, and the resulting repebody-mAP fusion protein was used for direct and sandwich immunoassays of hTNF-alpha. We demonstrate the utility and potential of the repebody-mAP fusion protein as an immuno-reagent by showing the sensitivity of 216 pg mL(-1) for hTNF-alpha in a sandwich immunoassay. Furthermore, this repebody-mAP fusion protein enabled the detection of hTNF-alpha spiked in a serum-supplemented medium with high accuracy and reproducibility. It is thus expected that a mAP-fused repebody can be broadly used as an immuno-reagent in immunoassays. (CB.V) 2016 Elsevier B.V. All rights reserved. | - |
dc.language | English | - |
dc.publisher | ELSEVIER SCIENCE BV | - |
dc.subject | NECROSIS-FACTOR-ALPHA | - |
dc.subject | SINGLE-DOMAIN ANTIBODY | - |
dc.subject | FUSION PROTEINS | - |
dc.subject | FACTOR RECEPTOR | - |
dc.subject | AFFINITY | - |
dc.subject | BINDER | - |
dc.subject | ASSAYS | - |
dc.subject | SCFV | - |
dc.title | Alkaline phosphatase-fused repebody as a new format of immuno-reagent for an immunoassay | - |
dc.type | Article | - |
dc.identifier.wosid | 000390629500021 | - |
dc.identifier.scopusid | 2-s2.0-84999885008 | - |
dc.type.rims | ART | - |
dc.citation.volume | 950 | - |
dc.citation.beginningpage | 184 | - |
dc.citation.endingpage | 191 | - |
dc.citation.publicationname | ANALYTICA CHIMICA ACTA | - |
dc.identifier.doi | 10.1016/j.aca.2016.11.013 | - |
dc.contributor.localauthor | Kim, Hak-Sung | - |
dc.contributor.nonIdAuthor | Kim, Yu Jung | - |
dc.contributor.nonIdAuthor | Hantschel, Oliver | - |
dc.contributor.nonIdAuthor | Lee, Seung-Goo | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Monomeric alkaline phosphatase | - |
dc.subject.keywordAuthor | Repebody | - |
dc.subject.keywordAuthor | Genetic fusion | - |
dc.subject.keywordAuthor | Immuno-reagent | - |
dc.subject.keywordAuthor | Immunoassay | - |
dc.subject.keywordPlus | NECROSIS-FACTOR-ALPHA | - |
dc.subject.keywordPlus | SINGLE-DOMAIN ANTIBODY | - |
dc.subject.keywordPlus | FUSION PROTEINS | - |
dc.subject.keywordPlus | FACTOR RECEPTOR | - |
dc.subject.keywordPlus | AFFINITY | - |
dc.subject.keywordPlus | BINDER | - |
dc.subject.keywordPlus | ASSAYS | - |
dc.subject.keywordPlus | SCFV | - |
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