Gold nanoparticle-based colorimetric assays for coagulation-related proteins and their inhibition reactions

Chen, Yi-You; Tseng, Chao-Wei; Chang, Hou‐Tai; Hung, Yu-Lun; Huang, Chih‐Ching

doi:10.1016/j.bios.2010.12.019

Cited by 32 publications

(8 citation statements)

References 51 publications

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“…In the above section, we have discussed GNP-based colorimetric assays for small molecules; similar detection systems have also been proposed for macromolecules such as proteins and whole cells (Table 1). Among several molecules that have been tested with GNP-based colorimetric strategies, thrombin is one of the predominant targets (Pavlov et al, 2004;Lu and Liu, 2006;Wei et al, 2007;Jian and Huang, 2011;Chen et al, 2012;Peng et al, 2013). Pavlov et al (2004) reported the use of an aptamerfunctionalized GNP, using thrombin as the target molecule, with a detection sensitivity limit of 20 nM.…”

Section: Detection Of Macromoleculesmentioning

confidence: 99%

Colorimetric detection of controlled assembly and disassembly of aptamers on unmodified gold nanoparticles

Gopinath

Lakshmipriya

Awazu

2014

Biosensors and Bioelectronics

216

128

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Aptamers are nucleic acid ligands that are generated artificially by in vitro selection and behave similar to antibodies. The development of aptamer-based sensing systems or strategies has been in vogue for the past few decades, because aptamers are smaller in size, stable, cheaper and undergo easier modifications. Owing to these advantages, several facile aptamer-based colorimetric strategies have been created by controlling the assembly and disassembly of aptamers on unmodified gold nanoparticle probes. As these kinds of assay systems are rapid and can be visualized unaided by instruments, they have recently become an attractive method of choice. The formation of purple-colored aggregates (attraction) from the red dispersed (repulsion) state of GNPs in the presence of mono- or divalent ions is the key principle behind this assay. Due to its simplicity and versatility, this assay can be an alternative to existing diagnostic assays. Here, we have investigated the critical elements involved in colorimetric assays, and have screened different proteins and small ligands to evaluate biofouling on GNPs.

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Section: Detection Of Macromoleculesmentioning

confidence: 99%

Colorimetric detection of controlled assembly and disassembly of aptamers on unmodified gold nanoparticles

Gopinath

Lakshmipriya

Awazu

2014

Biosensors and Bioelectronics

216

128

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“…55 because these two aptamers bind with the fibrinogen-binding exosite I. 65 We observed that the thrombin activity (1.0 nM) was inhibited to approximately 95% by TBA 15 (100 nM) and to approximately 60% by TBA 24 (100 nM), whereas TBA 29 (100 nM) exhibited no obvious anticoagulant activity. We also noted that hirudin, a 65-amino-acid polypeptide suppressed thrombin activity to ca.…”

Section: Analysis Of Thrombin-inducedmentioning

confidence: 93%

Monitoring Thrombin Generation and Screening Anticoagulants through Pulse Laser-Induced Fragmentation of Biofunctional Nanogold on Cellulose Membranes

Chiu

Unnikrishnan

et al. 2014

ACS Appl. Mater. Interfaces

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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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“…[16][17][18] In a similar way, a colorimetric assay to detect fibrinogen in plasma via the fibrinogeninduced aggregation of thrombin-conjugated Au NPs Thr-Au NPs has been reported. [19] Water-soluble near infrared CuInS 2 quantum dots (QDs) capped by mercaptopropionic acid [20] have been also used to monitor thrombin activity. The reported assay is based on the fluorescence quenching of the fibrinogen-CuInS 2 QDs complex triggered by thrombin.…”

Section: Introductionmentioning

confidence: 99%

CdS quantum dots generated in-situ for fluorometric determination of thrombin activity

et al. 2019

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A method is presented for sensitive determination of thrombin activity. It is based on (a) the interaction between fibrinogen after activation with thrombin, and (b) an enzymatic amplification step consisting of in-situ growth of CdS quantum dots (QDs). Fibrinogen is immobilized on the surface of the wells of a microplate and then incubated with a mixture of biotinylated fibrinogen and thrombin. Thrombin activates immobilized fibrinogen and free biotinylated fibrinogen. This leads to the formation of insoluble biotinylated fibrin that remains bound on the surface of the wells. Afterwards, the samples are incubated with avidin-labeled alkaline phosphatase (ALP) which binds to biotinylated fibrin. ALP hydrolyzes the substrate p-nitrophenyl phosphate (pNPP) under formation of phosphate ions which stabilize CdS QDs that are grown in-situ from cadmium(II) and sulfide. The generation of fibrin is correlated with the activity of thrombin. Increased thrombin concentration results in increased fluorescence that can be measured at excitation/emission wavelengths of 300/510 nm. The introduction of such an amplification step (the enzyme-triggered growth of QDs) allows for the quantification of thrombin in the picomolar concentration range, with a linear response up to 2.5 pM and a detection limit of 0.05 pM. The method was applied to the determination of thrombin activity in human plasma and of the thrombin inhibitor argatroban.

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Gold nanoparticle-based colorimetric assays for coagulation-related proteins and their inhibition reactions

Cited by 32 publications

References 51 publications

Colorimetric detection of controlled assembly and disassembly of aptamers on unmodified gold nanoparticles

Colorimetric detection of controlled assembly and disassembly of aptamers on unmodified gold nanoparticles

Monitoring Thrombin Generation and Screening Anticoagulants through Pulse Laser-Induced Fragmentation of Biofunctional Nanogold on Cellulose Membranes

CdS quantum dots generated in-situ for fluorometric determination of thrombin activity

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