title: Fibrinolysis and thrombosis of fibrinogen-modified gold nanoparticles for detection of fibrinolytic-related proteins.
abstract: Fibrinolysis (plasmin-mediated cleavage of fibrin structures) is a process in which fibrin clots can be removed from blood vessels, allowing the return of normal vascular function. Although several methods have been developed to measure plasmin activity and plasminogen (the plasmin precursor) concentrations, they are only moderately sensitive and quantitative and require large amounts of reagents, limiting their applicability. We developed two simple, label-free homogeneous assays using gold nanoparticles (Au NPs) for detection of fibrinolysis-related proteins and their activator (urokinase that converts plasminogen to plasmin) and inhibitor (α2-plasmin inhibitor that inhibits plasmin and plasminogen bound to fibrin). We used a fibrinolysis-based sensor, based on plasmin-mediated cleavage of fibrinogen-modified Au NPs (Fib-Au NPs) leading to aggregation of Au NPs, to determine plasmin activity in a biological medium mimic solution. A combination of thrombin (Thr) and Fib-Au NPs allowed us to analyze plasmin activity and plasminogen concentrations in serum through Thr-induced agglutination of Fib-Au NPs. The limit of detection (LOD; S/N=3) of this sensor for plasmin in serum was 0.4 nM (ca. 1.7×10(-4) unit mL(-1)). These label-free assays offer several advantages over conventional assays, including allowing rapid and simple readings with the naked eye or measurement by UV-vis absorption spectroscopy.
authors.affiliation: Institute of Bioscience and Biotechnology, National Taiwan Ocean University, 2, Pei-Ning Road, Keelung 20224, Taiwan.
authors.forename: Jyun-Wei
authors.initials: JW
authors.lastname: Jian
authors.forename: Wei-Chane
authors.initials: WC
authors.lastname: Chiu
authors.forename: Huan-Tsung
authors.initials: HT
authors.lastname: Chang
authors.forename: Pang-Hung
authors.initials: PH
authors.lastname: Hsu
authors.forename: Chih-Ching
authors.initials: CC
authors.lastname: Huang
mesh_terms: D000328:Adult; D015374:Biosensing Techniques / Q000379:methods*; D005340:Fibrinogen / Q000737:chemistry* / Q000378:metabolism; D005341:Fibrinolysin / Q000032:analysis* / Q000378:metabolism; D005342:Fibrinolysis; D006046:Gold / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D053758:Nanoparticles / Q000737:chemistry*; D010958:Plasminogen / Q000032:analysis* / Q000378:metabolism; D013917:Thrombin / Q000737:chemistry / Q000378:metabolism; D013927:Thrombosis / Q000175:diagnosis / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis / Q000378:metabolism; D055815:Young Adult; D000979:alpha-2-Antiplasmin / Q000032:analysis / Q000378:metabolism
chemical_list: D000979:alpha-2-Antiplasmin; D006046:Gold; D005340:Fibrinogen; D010958:Plasminogen; D013917:Thrombin; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator