title: Nanostructured bioluminescent sensor for rapidly detecting thrombin.
abstract: Thrombin plays a key role in thrombosis and hemostasis. The abnormal level of thrombin in body fluids may lead to different diseases, such as rheumatoid arthritis, glomerulonephritis, etc. Detection of thrombin level in blood and/or urine is one of important methods for medical diagnosis. Here, a bioluminescent sensor is developed for non-invasively and rapidly detecting thrombin in urine. The sensor is assembled through conjugating gold nanoparticles (Au NPs) and a recombinant protein containing Renilla luciferase (pRluc) by a peptide, which is thrombin specific substrate. The luciferase-catalyzed bioluminescence can be quenched by peptide-conjugating Au NPs. In the presence of thrombin, the short peptide conjugating luciferase and Au NPs is digested and cut off, which results in the recovery of bioluminescence due to the release of luciferase from Au NPs. The bioluminescence intensity at 470 nm is observed, and increases with increasing concentration of thrombin. The bioluminescence intensity of this designed sensor is significantly recovered when the thrombin digestion time lasts for 10 min. In addition, a similar linear relationship between luminescence intensity and the concentration of thrombin is found in the range of 8 nM to 8 μM in both buffer and human urine spiked samples. The limit of detection is as low as 80 pM. It is anticipated that our nanosensor could be a promising tool for clinical diagnosis of thrombin in human urine.
authors.affiliation: Department of Chemical and Biochemical Engineering, Western University, London, Ontario, Canada N6A 5B9.
authors.forename: Longyan
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Surgery, Western University, London, Ontario, Canada.
authors.forename: Yige
authors.initials: Y
authors.lastname: Bao
authors.affiliation: Department of Surgery, Western University, London, Ontario, Canada.
authors.forename: John
authors.initials: J
authors.lastname: Denstedt
authors.affiliation: Department of Chemical and Biochemical Engineering, Western University, London, Ontario, Canada N6A 5B9. Electronic address: jzhang@eng.uwo.ca.
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D003199:Computer Systems; D004867:Equipment Design; D019544:Equipment Failure Analysis; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation*; D049410:Luciferases, Renilla / Q000737:chemistry; D008163:Luminescent Measurements / Q000295:instrumentation*; D008164:Luminescent Proteins / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D036103:Nanotechnology / Q000295:instrumentation*; D013917:Thrombin / Q000652:urine*; D016482:Urinalysis / Q000295:instrumentation
chemical_list: D008164:Luminescent Proteins; D049410:Luciferases, Renilla; D013917:Thrombin
keywords: Bioluminescence resonance energy transfer sensor; Gold nanoparticle; Luciferase; Thrombin