Aptamer‐Conjugated Nanoparticles Efficiently Control the Activity of Thrombin

Shiang, Yen-Chun; Huang, Chih‐Ching; Wang, Tze-Hsiang; Chien, Chun-Wei; Chang, Huan‐Tsung

doi:10.1002/adfm.201000642

Cited by 59 publications

(70 citation statements)

References 61 publications

(33 reference statements)

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“…They exhibited a high binding affinity (K d = 3.4 10 À12 m) toward thrombinover 100 times higher than that of monovalent TBA 29 , 10 000 times higher than that of TBA 15 , and at least 10 times higher than reported previously for fusion aptamers, dendritic aptamers, and TBA 29 -AuNPs. [9,10] They exhibited TCT 91 times longer than without inhibitor. Moreover, they antagonized the anticoagulant activity of cTBA 15 /cTBA 29 -AuNPs 20 times faster than did cTBA.…”

Section: Resultsmentioning

confidence: 99%

“…15-TBA 15 /TBA 29 / sulf-Gal-AuNPs have the highest anticoagulation potency, -GGT TGG TGT GGT TGG-3' TBA 29 5'-AGT CCG TGG TAG GGC AGG TTG GGG TGA CT-3' cTBA 15 5'-CCA ACC ACA CCA ACC-3' cTBA 29 5 www.chemeurj.org which is consistent with the known tendency for a higher local concentration of TBA ligands to enhance binding affinity toward thrombin. [10] Gel electrophoresis is a technique sensitive to oligonucleotide conformation in oligonucleotide-AuNPs conjugates. [14] Figure S5 in the Supporting Information shows a 1.5 % agarose gel that contains aptamermodified AuNPs samples.…”

Section: Resultsmentioning

confidence: 99%

“…[17] TBA and its complementary sequences (cTBA) are known to be effective drug/antidote pairs for thrombin. [10] We investigated the effects of two antidotes, cTBA molecules and cTBA-conjugated AuNPs (cTBA-AuNPs), on the anticoagulant properties of the 15-TBA 15 /TBA 29 /sulf-Gal-AuNPs. Clotting mixtures of thrombin (5.0 nm), fibrinogen (1.14 mm), and 15-TBA 15 / TBA 29 /sulf-Gal-AuNPs (2.5 nm) were treated separately with 500 nm cTBA 15 /cTBA 29 (1:1) and 10 nm cTBA 15 / cTBA 29 -AuNPs (around 50 cTBA 15 and 50 cTBA 29 molecules per AuNP).…”

Section: Thrombin Clotting Time and Stabilitymentioning

confidence: 99%

“…[9] Recently, TBA 29 -conjugated gold nanoparticles (TBA 29 -AuNPs) were reported to provide 82-fold improved anticoagulant potency through thrombin-mediated coagulation made possible by steric blocking and a high binding affinity toward thrombin in solutions that mimic biological plasma. [10] However, its lack of both flexibility and multivalent interactions limit improvements in its potency, and in plasma samples it exhibited a clotting time only 2 times longer than for TBA 15 .…”

Section: Introductionmentioning

confidence: 99%

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Highly Efficient Control of Thrombin Activity by Multivalent Nanoparticles

Hsu

Chang

Chen

et al. 2011

Chemistry A European J

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We have demonstrated that the incorporation of sulfated galactose acid (sulf-Gal) into thrombin-binding-aptamer (TBA)-conjugated gold nanoparticles (TBA-AuNPs) enables highly effective inhibition of thrombin activity toward fibrinogen. AuNP bioconjugates (TBA(15)/TBA(29)/sulf-Gal-AuNPs) were prepared from 13 nm AuNPs, 15-mer thrombin-binding aptamer (TBA(15)), 29-mer thrombin-binding aptamer (TBA(29)), and sulf-Gal. The numbers of TBA and sulf-Gal molecules per AuNP proved to have a strong impact on inhibitory potency. The best results were observed for 15-TBA(15)/TBA(29)/sulf-Gal-AuNPs (with 15 TBA(15) and 15 TBA(29) molecules per AuNP), which, because of their particularly flexible conformation and multivalency, exhibited ultrahigh binding affinity toward thrombin (K(d)=3.4×10(-12) M) and thus extremely high anticoagulant (inhibitory) potency. Compared to the case without inhibitors (the "normal" value), their measured thrombin clotting time (TCT) was 91 times longer, whereas for TBA(15) alone it was only 7.2 times longer. Their anticoagulant activity was suppressed by TBA-complementary-sequence (cTBA)-modified AuNPs (cTBA(15)/cTBA(29)-AuNPs) at a rate that was 20 times faster than that of free cTBA(15)/cTBA(29). Thus, easily prepared, low-cost, multivalent AuNPs show great potential for biomedical control of blood clotting.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Thrombin Clotting Time and Stabilitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Highly Efficient Control of Thrombin Activity by Multivalent Nanoparticles

Hsu

Chang

Chen

et al. 2011

Chemistry A European J

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“…[15] The coagulation protein thrombin plays many roles in the coagulation cascade, converting soluble fibrinogen into insoluble strands of fibrin, as well as catalyzing many other coagulation-related reactions. [16] However, although TBA 29 interacts strongly with thrombin (K d % 0.5 nm), it lacks the enzymatic inhibitory function required for thrombin-mediated coagulation because it interacts with thrombin exosite 2 rather than its fibrinogen-binding exosite 1.…”

Section: Introductionmentioning

confidence: 99%

Colorimetric Detection of DNA by Modulation of Thrombin Activity on Gold Nanoparticles

Jian

Huang

2011

Chemistry A European J

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A colorimetric, non-cross-linking aggregation-based gold-nanoparticle (AuNP) probe has been developed for the detection of DNA and the analysis of single-nucleotide polymorphism (SNP). The probe acts by modulating the enzyme activity of thrombin relative to fibrinogen. A thrombin-binding aptamer with a 29-base-long oligonucleotide (TBA(29)) assembled on the nanoparticles (TBA(29)-AuNPs) through sandwich DNA hybridization was found to possess ultra-high anticoagulant potency. The enzyme inhibition of thrombin was determined by thrombin-induced aggregation of fibrinogen-functionalized 56 nm AuNPs (Fib-AuNPs). The potency of the inhibition of TBA(29)-AuNPs relative to thrombin--and thus the degree of aggregation of the Fib-AuNPs--is highly dependent on the concentration of perfectly matched DNA (DNA(pm)). Under optimal conditions [Tris-HCl (20 mM, pH 7.4), KCl (5 mM), MgCl(2) (1 mM), CaCl(2) (1 mM), NaCl (150 mM), thrombin (10 pM), and TBA(29)-AuNPs (20 pM)], the new TBA(29)-AuNP/Fib-AuNP probe shows linear sensitivity to DNA(pm) in the concentration range 20-500 pM with a correlation coefficient of 0.96. The limit of detection for DNA(pm) was experimentally determined to be 12 pM, based on a signal-to-noise ratio (S/N) of 3. The new probe was successfully applied to the analysis of an SNP that is responsible for sickle cell anemia. Relative to conventional molecular-beacon-based probes, the new probe offers the advantages of higher sensitivity and selectivity towards DNA and lower cost, showing its great potential for practical studies of SNPs.

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Self‐Assembly of Antimicrobial Peptides on Gold Nanodots: Against Multidrug‐Resistant Bacteria and Wound‐Healing Application

Chen

Chang

et al. 2015

Adv Funct Materials

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271

151

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Photoluminescent gold nanodots (Au NDs) are prepared via etching and codeposition of hybridized ligands, an antimicrobial peptide (surfactin; SFT), and 1‐dodecanethiol (DT), on gold nanoparticles (≈3.2 nm). As‐prepared ultrasmall SFT/DT–Au NDs (size ≈2.5 nm) are a highly efficient antimicrobial agent. The photoluminescence properties and stability as well as the antimicrobial activity of SFT/DT–Au NDs are highly dependent on the density of SFT on Au NDs. Relative to SFT, SFT/DT–Au NDs exhibit greater antimicrobial activity, not only to nonmultidrug‐resistant bacteria but also to the multidrug‐resistant bacteria. The minimal inhibitory concentration values of SFT/DT–Au NDs are much lower (>80‐fold) than that of SFT. The antimicrobial activity of SFT/DT–Au NDs is mainly due to the synergistic effect of SFT and DT–Au NDs on the disruption of the bacterial membrane. In vitro cytotoxicity and hemolysis analyses have revealed superior biocompatibility of SFT/DT–Au NDs than that of SFT. Moreover, in vivo methicillin‐resistant S. aureus–infected wound healing studies in rats show faster healing, better epithelialization, and are more efficient in the production of collagen fibers when SFT/DT–Au NDs are used as a dressing material. This study suggests that the SFT/DT–Au NDs are a promising antimicrobial candidate for preclinical applications in treating wounds and skin infections.

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Aptamer‐Conjugated Nanoparticles Efficiently Control the Activity of Thrombin

Cited by 59 publications

References 61 publications

Highly Efficient Control of Thrombin Activity by Multivalent Nanoparticles

Highly Efficient Control of Thrombin Activity by Multivalent Nanoparticles

Colorimetric Detection of DNA by Modulation of Thrombin Activity on Gold Nanoparticles

Self‐Assembly of Antimicrobial Peptides on Gold Nanodots: Against Multidrug‐Resistant Bacteria and Wound‐Healing Application

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