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Monitoring thrombin generation and screening anticoagulants through pulse laser-induced fragmentation of biofunctional nanogold on cellulose membranes.

Authors
  • Li, Yu-Jia1
  • Chiu, Wei-Jane
  • Unnikrishnan, Binesh
  • Huang, Chih-Ching
  • 1 Institute of Bioscience and Biotechnology, National Taiwan Ocean University , Keelung 20224, Taiwan. , (Taiwan)
Type
Published Article
Journal
ACS Applied Materials & Interfaces
Publisher
American Chemical Society
Publication Date
Sep 10, 2014
Volume
6
Issue
17
Pages
15253–15261
Identifiers
DOI: 10.1021/am503615c
PMID: 25141032
Source
Medline
Keywords
License
Unknown

Abstract

Thrombin generation (TG) has an important part in the blood coagulation system, and monitoring TG is useful for diagnosing various health issues related to hypo-coagulability and hyper-coagulability. In this study, we constructed probes by using mixed cellulose ester membranes (MCEMs) modified with gold nanoparticles (Au NPs) for monitoring thrombin activity using laser desorption/ionization mass spectrometry (LDI-MS). The LDI process produced Au cationic clusters ([Au(n)](+); n = 1-3) that we detected through MS. When thrombin reacted with fibrinogen on the Au NPs-MCEMs, insoluble fibrin was formed, hindering the formation of Au cationic clusters and, thereby, decreasing the intensity of their signals in the mass spectrum. Accordingly, we incorporated fibrinogen onto the Au NPs-MCEMs to form Fib-Au NPs-MCEM probes to monitor TG with good selectivity (>1000-fold toward thrombin with respect to other proteins or enzymes) and sensitivity (limit of detection for thrombin of ca. 2.5 pM in human plasma samples). Our probe exhibited remarkable performance in monitoring the inhibition of thrombin activity by direct thrombin inhibitors. Analyses of real samples using our new membrane-based probe suggested that it will be highly useful in practical applications for the effective management of hemostatic complications.

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