Generation of a Recombinant scFv against Deoxycholic Acid and Its Conversion to a Quenchbody for One-Step Immunoassay-Reference-Cited by-同舟云学术

Generation of a Recombinant scFv against Deoxycholic Acid and Its Conversion to a Quenchbody for One-Step Immunoassay

Published:2023-09-30 Issue:5 Volume:6 Page:90
ISSN:2409-9279
Container-title:Methods and Protocols
language:en
Short-container-title:MPs

Author:

Ueda Hiroshi¹^ORCID,Jeong Hee-Jin²^ORCID

Affiliation:

1. Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, Yokohama 226-8503, Japan

2. Department of Biological and Chemical Engineering, Hongik University, Sejong 30016, Republic of Korea

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.

Funder

National Research Foundation of Korea (NRF) grant funded by the Korean government

Publisher

MDPI AG

Subject

Biochemistry, Genetics and Molecular Biology (miscellaneous),Structural Biology,Biotechnology

Link

https://www.mdpi.com/2409-9279/6/5/90/pdf

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