Generation of Q-bead against bone Gla protein with simplified preparation steps
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, J.-P. | - |
dc.contributor.author | Kim, S.-H. | - |
dc.contributor.author | Ueda, H. | - |
dc.contributor.author | Jeong, H.-J. | - |
dc.date.accessioned | 2023-04-24T03:40:32Z | - |
dc.date.available | 2023-04-24T03:40:32Z | - |
dc.date.issued | 2023-05-01 | - |
dc.identifier.issn | 0022-1759 | - |
dc.identifier.issn | 1872-7905 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/31102 | - |
dc.description.abstract | Quenchbody (Q-body)-based immunoassays enable the detection of antigen within a few minutes with high sensitivity and specificity, thereby revealing their applicability as biosensors for quantifying several biomolecules of interest; however, while producing a Q-body, it is necessary to eliminate the unconjugated dye after labeling to separate the Q-body from the capturing bead and to change the buffer using ultrafiltration, which is time-consuming and leads to yield reduction. In this study, we generated a recombinant single chain variable fragment against bone Gla protein as a model antibody. We labeled the antibody with a dye to generate a Q-body and subsequently added affinity beads to the Q-body mixture. After washing, we directly added antigen without extracting the Q-body from the bead and then measured the fluorescence intensity. As a result, the antigen-dependent fluorescence response was obtained from “Q-bead”, which was almost the same as that of the Q-body generated according to the conventional method. The Q-bead was generated within only 2.5 h, thus requiring an hour and two steps less than those required for the generation of the traditional Q-body. No expensive Flag peptide was required to recover the total antibody from beads. Moreover, the ultra-filtration step was eliminated in this bead-based method, leading to improved convenience and cost- and time-saving attributes. The Q-bead-based assay can be used as a standard protocol for simple and rapid analysis of antibody-based molecular detection. © 2023 Elsevier B.V. | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | Elsevier B.V. | - |
dc.title | Generation of Q-bead against bone Gla protein with simplified preparation steps | - |
dc.type | Article | - |
dc.publisher.location | 네델란드 | - |
dc.identifier.doi | 10.1016/j.jim.2023.113471 | - |
dc.identifier.scopusid | 2-s2.0-85152109106 | - |
dc.identifier.wosid | 001029658100001 | - |
dc.identifier.bibliographicCitation | Journal of Immunological Methods, v.516 | - |
dc.citation.title | Journal of Immunological Methods | - |
dc.citation.volume | 516 | - |
dc.type.docType | Article | - |
dc.description.isOpenAccess | N | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Biochemistry & Molecular Biology | - |
dc.relation.journalResearchArea | Immunology | - |
dc.relation.journalWebOfScienceCategory | Biochemical Research Methods | - |
dc.relation.journalWebOfScienceCategory | Immunology | - |
dc.subject.keywordAuthor | Bone Gla protein | - |
dc.subject.keywordAuthor | Fluorescence sensor | - |
dc.subject.keywordAuthor | On-bead assay | - |
dc.subject.keywordAuthor | One-pot immunoassay | - |
dc.subject.keywordAuthor | Quenchbody | - |
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