A Bioluminescent Probe for Salivary Cortisol

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
DOI: 10.1021/bc200220k Publication Date: 2011-08-22T12:09:34Z
ABSTRACT
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.
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