Generation of a Recombinant scFv against Deoxycholic Acid and Its Conversion to a Quenchbody for One-Step Immunoassay.
Hiroshi UedaHee-Jin JeongPublished in: Methods and protocols (2023)
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.
Keyphrases
- loop mediated isothermal amplification
- label free
- sensitive detection
- escherichia coli
- papillary thyroid
- oxidative stress
- real time pcr
- quantum dots
- cell free
- photodynamic therapy
- squamous cell carcinoma
- squamous cell
- young adults
- living cells
- lymph node metastasis
- cystic fibrosis
- klebsiella pneumoniae
- single molecule
- pet ct