A proof of concept application of aptachain: ligand-induced self-assembly of a DNA aptamer

Neves, Miguel A. D.; Slavkovic, Sladjana; Reinstein, Oren; Shoara, Aron A.; Johnson, Phyllis E.

doi:10.1039/c8ra07462c

Cited by 14 publications

(5 citation statements)

References 34 publications

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“…The first DNA fragment is used as a recognition element, while the other for the electrochemical signal generation. The split‐aptamer used here can detect a small molecule, represented by adenosine; besides, the increasing development of oligonucleotide engineering allows replication of this strategy to other small molecules of toxicological or metabolic relevance . The assay achieves state‐of‐the‐art figures of merit as it can be observed in Table .…”

Section: Resultsmentioning

confidence: 99%

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Redox Polyelectrolyte Modified Gold Nanoparticles Enhance the Detection of Adenosine in an Electrochemical Split‐Aptamer Assay

et al. 2020

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We present an electrochemical sandwich‐type assay based on the splitting of an aptamer into two fragments. Gold nanoparticles are modified with one of the fragments and a redox polyelectrolyte. The first is used as the recognition element, while the other for the electrochemical signal generation. The split‐aptamer used here can detect adenosine, used as a model system for recognizing small molecules. The multiple binding sites on the nanoparticle, along with the high number of redox probes, yield a selective and sensitive assay for adenosine, achieving a limit of detection of 3.1 nM and a linear range up to 75 nM. The obtained results are analyzed in terms of the nanoparticle and electrode architectures. The assay can be easily extended to other small molecules and sandwich assays, representing a promising tool for detecting metabolites at the nanomolar level.

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Section: Resultsmentioning

confidence: 99%

“…In recent years, the sequence of several aptamers has been split into two parts, maintaining their affinity, allowing the detection of a species in a similar way than the antibody sandwich assay. In a sandwich assay using electrochemical signaling, the capture antibody is immobilized on the working electrode surface and binds the analyte.…”

Section: Introductionmentioning

confidence: 99%

Redox Polyelectrolyte Modified Gold Nanoparticles Enhance the Detection of Adenosine in an Electrochemical Split‐Aptamer Assay

et al. 2020

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“…The use of the hybridization chain reaction (HCR) forming such self-assembled 1D DNA chains for signal amplification has been widely developed in recent years [34][35][36][37][38][39][40]. However, its use with aptamer recognition to form what are called aptachains, is still in its infancy [3,[41][42][43][44]. In our case, the adenosine target served as the trigger for thermodynamic control of the linear DNA nanostructure formation in solution.…”

Section: Introductionmentioning

confidence: 99%

Melting Curve Analysis of Aptachains: Adenosine Detection with Internal Calibration

Saint-Pierre

Gasparutto

et al. 2021

Biosensors

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Small molecules are ubiquitous in nature and their detection is relevant in various domains. However, due to their size, sensitive and selective probes are difficult to select and the detection methods are generally indirect. In this study, we introduced the use of melting curve analysis of aptachains based on split-aptamers for the detection of adenosine. Aptamers, short oligonucleotides, are known to be particularly efficient probes compared to antibodies thanks to their advantageous probe/target size ratio. Aptachains are formed from dimers with dangling ends followed by the split-aptamer binding triggered by the presence of the target. The high melting temperature of the dimers served as a calibration for the detection/quantification of the target based on the height and/or temperature shift of the aptachain melting peak.

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“…Organosilane based SAMs are ordered molecular assemblies, comprised of linking molecules, formed by spontaneous covalent attachment onto many hydroxyl (─OH) terminated substrates (Figure 1a). [3][4][5] They are commonly utilized in applications across many scientific and engineering disciplines such as the development of fouling resistant surfaces, [4][5][6] improved substrate biocompatibility, [6,7] biosensing, [8,9] and cellular signaling and interactions. [10] Literature reports of the covalent immobilization of monomeric proteins and nucleic acids are quite common, however, there are no literature reports of the sitedirected, oriented, covalent coupling of heterodimeric proteins to organic SAMs.…”

Section: Introductionmentioning

confidence: 99%

Heterodimeric Protein Surface‐Coupling Platform: Immobilization of Conformation Switchable and Functional αIIbβ3 Integrin

Neves,

Slavkovic,

Wang

et al. 2023

Adv Materials Inter

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Many biotechnologies require direct contact between biomolecules and (semi)solid substrates. However, little information is available regarding site‐directed covalent surface immobilization of heterodimeric proteins. Integrins are heterodimeric, conformationally dependent membrane adhesion‐receptors, which are important in the (patho)biology of almost all human diseases. In this study, a biomimetic‐platform is developed for covalent and site‐directed immobilization of oriented integrin heterodimers onto most hydroxyl bearing surfaces, herein demonstrated on both glass‐slides and silica‐particles. This platform consists of a self‐assembled monolayer, upon which C‐terminal modified αIIbβ3 integrin extracellular domains are oriented and covalently anchored. As with cell surface integrins, the conformation of immobilized αIIbβ3 is switchable and can be modulated to the active ligand‐binding conformation by divalent cations. Furthermore, the αIIbβ3‐coupled silica particles display platelet‐mimetic hemostat function and co‐aggregate with platelets from both wild‐type and fibrinogen/von Willebrand factor double deficient mice, facilitating αIIbβ3‐non‐classical ligand discoveries. This work provides a biomaterial platform for functional multimeric protein‐substrate coupling, which should have broad impact on multiple fields of biology, biotechnology, and clinical diagnosis/therapy.

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A proof of concept application of aptachain: ligand-induced self-assembly of a DNA aptamer

Abstract: We present the concept of aptachain. An aptamer is split into two overlapping strands that form an oligomer when it binds its target. Aptachain formation can be used to detect ligand binding and may be beneficial in other biotechnology applications.

Cited by 14 publications

References 34 publications

Redox Polyelectrolyte Modified Gold Nanoparticles Enhance the Detection of Adenosine in an Electrochemical Split‐Aptamer Assay

Redox Polyelectrolyte Modified Gold Nanoparticles Enhance the Detection of Adenosine in an Electrochemical Split‐Aptamer Assay

Melting Curve Analysis of Aptachains: Adenosine Detection with Internal Calibration

Heterodimeric Protein Surface‐Coupling Platform: Immobilization of Conformation Switchable and Functional αIIbβ3 Integrin

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