Different strategies to develop an electrochemical thrombin aptasensor

Mir, Mònica; Vreeke, Mark; Katakis, Ioanis

doi:10.1016/j.elecom.2005.12.022

Cited by 155 publications

(109 citation statements)

References 33 publications

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“…The use of chromogenic substrate in TLAA also makes colorimetric assay feasible as the generated product is colored, and target detection can be achieved by observing the color change of the solution without using instruments, which will be useful for point-of-care applications. Because the generated product can be measured with electrochemical method [26,27], it is also possible to apply electrochemical detection method for TLAA. Though a sandwich format TLAA is demonstrated here for PDGF-BB, other assay formats of TLAA (e.g., competitive format) can be expected.…”

Section: Performance Of Assaysmentioning

confidence: 99%

“…Therefore, several highly sensitive assays for thrombin have used aptamer binding to one site and the thrombin enzyme activity on the other site. For example, thrombin could be captured on a solid support, and captured thrombin could subsequently catalyze the hydrolysis of small peptide substrates [26][27][28][29][30][31]. We have achieved sensitive detection of thrombin by taking advantage of affinity preconcentration of thrombin onto aptamercoated magnetic beads [29][30][31].…”

Section: Introductionmentioning

confidence: 99%

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Thrombin-linked aptamer assay for detection of platelet derived growth factor BB on magnetic beads in a sandwich format

Guo

Zhao

2016

Talanta

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a b s t r a c tHere we describe a thrombin-linked aptamer assay (TLAA) for protein by using thrombin as an enzyme label, harnessing enzyme activity of thrombin and aptamer affinity binding. TLAA converts detection of specific target proteins to the detection of thrombin by using a DNA sequence that consists of two aptamers with the first aptamer binding to the specific target protein and the second aptamer binding to thrombin. Through the affinity binding, the thrombin enzyme is labeled on the protein target, and thrombin catalyzes the hydrolysis of small peptide substrate into product, generating signals for quantification. As a proof of principle, we show a sandwich TLAA for platelet derived growth factor BB (PDGF-BB) by using anti-PDGF-BB antibody coated on magnetic beads and an oligonucleotide containing the aptamer for PDGF-BB and the aptamer for thrombin. The binding of PDGF-BB to both the antibody and the aptamer results in labeling the complex with thrombin. We achieved detection of PDGF-BB at 16 pM. This TLAA contributes a new application of thrombin and its aptamer in bioanalysis, and shows potentials in assay developments.

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Section: Performance Of Assaysmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Thrombin-linked aptamer assay for detection of platelet derived growth factor BB on magnetic beads in a sandwich format

Guo

Zhao

2016

Talanta

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“…In a simple approach, TBA was non-specifically immobilized on the electrode surface, and thrombin was detected using the HRP label, allowing a 3.5 nM detection limit, sufficient for clinical diagnostic of metastatic lung cancer, where the concentration of thrombin level detected was 5.4 nM [56].…”

Section: Structure-switching G4 Electrochemical Aptasensormentioning

confidence: 99%

“…The first aptamer was generally immobilized onto gold via a thiol [56][57][58][59][60] and, more recently, magnetic beads [61,62]. The labels of the second aptamer were either redox molecules, nanocomposites [60], nanoparticles [57,63], quantum dots [58,59] or enzymes, with catalytic activity that transformed the substrate into an electroactive product [56,64,65].…”

Section: Sandwich-type G4 Electrochemical Aptasensormentioning

confidence: 99%

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Guanine Quadruplex Electrochemical Aptasensors

Chiorcea-Paquim

Oliveira-Brett

2016

Chemosensors

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Guanine-rich nucleic acids are able to self-assemble into G-quadruplex four-stranded secondary structures, which are found at the level of telomeric regions of chromosomes, oncogene promoter sequences and other biologically-relevant regions of the genome. Due to their extraordinary stiffness and biological role, G-quadruples become relevant in areas ranging from structural biology to medicinal chemistry, supra-molecular chemistry, nanotechnology and biosensor technology. In addition to classical methodologies, such as circular dichroism, nuclear magnetic resonance or crystallography, electrochemical methods have been successfully used for the rapid detection of the conformational changes from single-strand to G-quadruplex. This review presents recent advances on the G-quadruplex electrochemical characterization and on the design and applications of G-quadruplex electrochemical biosensors, with special emphasis on the G-quadruplex aptasensors and hemin/G-quadruplex peroxidase-mimicking DNAzyme biosensors.

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Electrochemical Detection of Proteins

Bartošík

Doneux

Pechan

et al. 2009

Encyclopedia of Analytical Chemistry

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The advantages of electrochemical methods, such as low cost, rapidity, and sensitivity, make these methods very promising in the booming fields of proteomics and biomedicine. Electroanalysis of proteins relies on redox properties of nonprotein centers within conjugated proteins, application of various labels attached to the protein molecule, or label‐free detection of nonconjugated proteins based on their intrinsic electroactivity. Until recently, electrochemistry of proteins was mainly focused on a relatively small group of conjugated proteins, yielding reversible electrochemistry. This article discusses the new methods that were developed to detect and analyze practically all proteins. Different strategies for electrochemical detection of proteins are presented. Electrode modification by biomolecular receptors (antibodies, aptamers) and their potential use for detection of specific proteins, e.g. disease markers, can be utilized in the development of various sensors. Label‐free electrochemical study of nonconjugated proteins, involving impedance measurements at various electrodes, faradaic signals of tyrosine and tryptophan residues at carbon electrodes and cysteine residues, and electrocatalytic signals at mercury and solid amalgam electrodes have been used. Among them, well‐developed chronopotentiometric peak (peak H) due to catalytic hydrogen evolution shows remarkable sensitivity to local and global changes in protein structure. Recent developments in protein electrochemistry open the way for application of electrochemical analysis in proteomics and biomedicine.

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Different strategies to develop an electrochemical thrombin aptasensor

Cited by 155 publications

References 33 publications

Thrombin-linked aptamer assay for detection of platelet derived growth factor BB on magnetic beads in a sandwich format

Thrombin-linked aptamer assay for detection of platelet derived growth factor BB on magnetic beads in a sandwich format

Guanine Quadruplex Electrochemical Aptasensors

Electrochemical Detection of Proteins

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