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Generation of a Recombinant scFv against Deoxycholic Acid and Its Conversion to a Quenchbody for One-Step Immunoassayopen access
- Authors
- Ueda, Hiroshi; Jeong, Hee-Jin
- Issue Date
- Oct-2023
- Publisher
- Multidisciplinary Digital Publishing Institute (MDPI)
- Keywords
- deoxycholic acid; hapten; immunoassay; Quenchbody; recombinant antibody; single-chain variable fragment
- Citation
- Methods and Protocols, v.6, no.5
- Journal Title
- Methods and Protocols
- Volume
- 6
- Number
- 5
- ISSN
- 2409-9279
2409-9279
- Abstract
- Development of a rapid detection method for deoxycholic acid (DCA) is crucial for its diagnosis in the early stages of inflammation and cancer. In this study, we expressed a soluble recombinant anti-DCA single-chain variable fragment (scFv) in Escherichia coli. To convert scFv into a Quenchbody (Q-body), we labeled scFv using commercially available maleimide-linked fluorophores. The TAMRA-C5-maleimide-conjugated Q-body showed the highest response within a few minutes of DCA addition, indicating its applicability as a wash-free immunoassay probe for onsite DCA detection. © 2023 by the authors.
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Related Researcher
- Jeong, Hee-Jin
- Science & Technology (Biological and Chemical Engineering)