Functional gold nanomaterials for the inhibition of the activity of thrombin and for the detection of ATP
Date Issued
2012
Date
2012
Author(s)
Liao, Yu-Ju
Abstract
There are two parts in this thesis: the first part is focused on that we prepared thrombin-binding aptamer-conjugated gold nanoparticles (TBA−Au NPs) through a molecularly imprinted (MP) approach, which provided highly efficient inhibition activity toward the polymerization of fibrinogen; another is to prepared actin-conjugated gold-silver nanorods (Ac−Au/Ag NRs) to measure Adenosine-5''-triphosphate (ATP) concentration in the aqueous solution by colorimetric method. MP-TBA15/TBA29-T15−Au NPs provided the highest binding affinity toward thrombin with a dissociation constant of 5.2 × 10−11 M. Finally, we tested the thrombin clotting time (TCT) measurements in plasma samples and found that MP-TBA15/TBA29-T15−Au NPs had greater anticoagulation activity than heparin, argatroban, hirudin and warfarin. In addition, we demonstrated that thrombin induced the formation of aggregates from MP-TBA15-T15−Au NPs and MP-TBA29-T15−Au NPs, thereby allowing the colorimetric detection of thrombin at the nanomolar level in serum samples. The second part is to demonstrated a colorimetric sensing approach for the determination of the concentration of adenosine-5''-triphosphate (ATP) using actin-conjugated gold nanorods (Ac−Au/Ag NRs). In the presence of the ATP, the color of the Ac−Au/Ag NRs solution changed from red to purple as a result of actin polymerization. For mixtures of ATP, polymerization buffer (40.0 mM NaCl and 2.0 mM MgCl2) and physiological buffer (25 mM Tris-HCl (pH 7.4), 150 mM NaCl, 5.0 mM KCl, 1.0 mM MgCl2, 1.0 mM CaCl2), a linear correlation (R2=0.99) existed between the ratio of the extinctions of the Ac−Au NRs at 900 and 730 nm (Ex900/730) and the concentration of ATP.
Subjects
gold nanomaterial
aptamer
molecularly imprinted technology
thrombin
ATP
Type
thesis
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