One-step detection of procollagen type III N-terminal peptide as a fibrosis biomarker using fluorescent immunosensor (quenchbody)
DC Field | Value | Language |
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dc.contributor.author | Yi, Joon-Yeop | - |
dc.contributor.author | Ryu, Jaewon | - |
dc.contributor.author | Jeong, Yujin | - |
dc.contributor.author | Cho, Yoeseph | - |
dc.contributor.author | Kim, Minyoung | - |
dc.contributor.author | Jeon, Mijin | - |
dc.contributor.author | Park, Hee Ho | - |
dc.contributor.author | Hwang, Nathaniel S. | - |
dc.contributor.author | Jeong, Hee-Jin | - |
dc.contributor.author | Sung, Changmin | - |
dc.date.accessioned | 2024-07-22T02:30:20Z | - |
dc.date.available | 2024-07-22T02:30:20Z | - |
dc.date.issued | 2024-08-15 | - |
dc.identifier.issn | 0003-2670 | - |
dc.identifier.issn | 1873-4324 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/hongik/handle/2020.sw.hongik/33372 | - |
dc.description.abstract | Background: Procollagen type III N-terminal peptide (P-III-NP) is a fibrosis biomarker associated with liver and cardiac fibrosis. Despite the value of P-III-NP as a biomarker, its analysis currently relies on enzyme-linked immunosorbent assays (ELISA) and radioimmunoassays (RIA), which require more than 3 h. To facilitate early diagnosis and treatment through rapid biomarker testing, we developed a one-step immunoassay for P-III-NP using a quenchbody, which is a fluorescence-labeled immunosensor for immediate signal generation. Results: To create quenchbodies, the total mRNA of P-III-NP antibodies was extracted from early-developed hybridoma cells, and genes of variable regions were obtained through cDNA synthesis, inverse PCR, and sequencing. A single-chain variable fragment (scFv) with an N-terminal Cys-tag was expressed in E. coli Shuffle T7, resulting in a final yield of 9.8 mg L−1. The fluorescent dye was labeled on the Cys-tag of the anti-P-III-NP scFv using maleimide-thiol click chemistry, and the spacer arm lengths between the maleimide-fluorescent dyes were compared. Consequently, a TAMRA-C6-labeled quenchbody exhibited antigen-dependent fluorescence signals and demonstrated its ability to detect P-III-NP at concentrations as low as 0.46 ng mL−1 for buffer samples, 1.0 ng mL−1 for 2 % human serum samples. Significance: This one-step P-III-NP detection method provides both qualitative and quantitative outcomes within a concise 5-min timeframe. Furthermore, its application can be expanded using a 96-well platform and human serum, making it a high-throughput and sensitive method for testing fibrotic biomarkers. © 2024 The Author(s) | - |
dc.language | 영어 | - |
dc.language.iso | ENG | - |
dc.publisher | Elsevier B.V. | - |
dc.title | One-step detection of procollagen type III N-terminal peptide as a fibrosis biomarker using fluorescent immunosensor (quenchbody) | - |
dc.type | Article | - |
dc.publisher.location | 네델란드 | - |
dc.identifier.doi | 10.1016/j.aca.2024.342887 | - |
dc.identifier.scopusid | 2-s2.0-85196410777 | - |
dc.identifier.bibliographicCitation | Analytica Chimica Acta, v.1317 | - |
dc.citation.title | Analytica Chimica Acta | - |
dc.citation.volume | 1317 | - |
dc.type.docType | Article | - |
dc.description.isOpenAccess | Y | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.subject.keywordAuthor | Fibrosis biomarker | - |
dc.subject.keywordAuthor | fluorescence | - |
dc.subject.keywordAuthor | Immunosensor | - |
dc.subject.keywordAuthor | Procollagen type III N-Terminal peptide | - |
dc.subject.keywordAuthor | Quenchbody | - |
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