A Bioluminescent Probe for Salivary Cortisol

• DOI: 10.1021/bc200220k
• Publication Date: 2011-08-22T12:09:34Z
• Authors: Sung Bae Kim; Yasuhiro Takenaka; Masaki Torimura
• Keywords: 0303 health sciences; Binding Sites; Hydrocortisone; Enzyme-Linked Immunosorbent Assay; Sensitivity and Specificity; 3. Good health; 03 medical and health sciences; Receptors, Glucocorticoid; Genes, Reporter; COS Cells; Chlorocebus aethiops; Luminescent Measurements; Animals; Humans; Luciferases; Promoter Regions, Genetic; Saliva; Glucocorticoids

Cortisol is a classical biomarker for the stress levels of human beings. We fabricated highly sensitive bioluminescent probes salivary cortisol. The following strategies were contrived in molecular design. Gaussia princeps luciferase (GLuc) was dissected into two fragments, between which an N-terminal-extended ligand binding domain glucocorticoid receptor (GR HLBD), named Simgr4, inserted. First, this unique single-chain probe then situated downstream response element (GRE) promoter reporter-gene system constructing ON–OFF switches Second, circularly permutated (CP) variant Simgr4 formulated. exerted improved signal-to-background (S/B) ratio 8.5 to Furthermore, 10× enhanced detection limit cortisol and long dynamic range from 10–9 10–6 M cortisol, covering all normal clinical ranges serum, urine, saliva. This optimized successfully determined daily fluctuations correlations with those by ELISA. study first investigate contribution HLBD nuclear multiple cortisols.