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Download fileMonitoring Thrombin Generation and Screening Anticoagulants through Pulse Laser-Induced Fragmentation of Biofunctional Nanogold on Cellulose Membranes
journal contribution
posted on 2014-09-10, 00:00 authored by Yu-Jia Li, Wei-Jane Chiu, Binesh Unnikrishnan, Chih-Ching HuangThrombin 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 ([Aun]+; 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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Keywords
probeblood coagulation systemcellulose ester membranesmass spectrummonitoring thrombin activitymonitoring TGMonitoring Thrombin GenerationBiofunctional Nanogoldhealth issuesthrombin inhibitorsMSNP2.5 pMcationic clustersthrombin activityLDI processhemostatic complicationsScreening Anticoagulantsgold nanoparticlesCellulose MembranesThrombin generationMCEMplasma samples