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A Nucleic Acid Nanostructure Built through On-Electrode Ligation for Electrochemical Detection of a Broad Range of Analytes.

ABSTRACT: For an assay to be most effective in point-of-care clinical analysis, it needs to be economical, simple, generalizable, and free from tedious workflows. While electrochemistry-based DNA sensors reduce instrumental costs and eliminate complicated procedures, there remains a need to address probe costs and generalizability, as numerous probes with multiple conjugations are needed to quantify a wide range of biomarkers. In this work, we have opened a route to circumvent complicated multiconjugation schemes using enzyme-catalyzed probe construction directly on the surface of the electrode. With this, we have created a versatile DNA nanostructure probe and validated its effectiveness by quantification of proteins (streptavidin, anti-digoxigenin, anti-tacrolimus) and small molecules (biotin, digoxigenin, tacrolimus) using the same platform. Tacrolimus, a widely prescribed immunosuppressant drug for organ transplant patients, was directly quantified with electrochemistry for the first time, with the assay range matching the therapeutic index range. Finally, the stability and sensitivity of the probe was confirmed in a background of minimally diluted human serum.

SUBMITTER: Somasundaram S

PROVIDER: S-EPMC6800171 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature

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A Nucleic Acid Nanostructure Built through On-Electrode Ligation for Electrochemical Detection of a Broad Range of Analytes.

Somasundaram Subramaniam S Easley Christopher J CJ

Journal of the American Chemical Society 20190709 29

For an assay to be most effective in point-of-care clinical analysis, it needs to be economical, simple, generalizable, and free from tedious workflows. While electrochemistry-based DNA sensors reduce instrumental costs and eliminate complicated procedures, there remains a need to address probe costs and generalizability, as numerous probes with multiple conjugations are needed to quantify a wide range of biomarkers. In this work, we have opened a route to circumvent complicated multiconjugation ...[more]

PMID: 31257869

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Project description:Electrodeposition is an electrochemical method employed to deposit stable and robust gold nanoparticles (AuNPs) on electrode surfaces for creating chemically modified electrodes (CMEs). The use of several electrodeposition techniques with different experimental parameters allow in obtaining various surface morphologies of AuNPs deposited on the electrode surface. By considering the electrodeposition of AuNPs in various background electrolytes could play an important strategy in finding the most suitable formation of the electrodeposited AuNP films on the electrode surface. This is because different electrode roughnesses can have different effects on the electrochemical activities of the modified electrodes. Thus, in this study, the electrodeposition of AuNPs onto the glassy carbon (GC) electrode surfaces in various aqueous neutral and acidic electrolytes was achieved by using the cyclic voltammetry (CV) technique with no adjustable CV parameters. Then, surface morphologies and electrochemical activities of the electrodeposited AuNPs were investigated using scanning electron microscopy (SEM), atomic force microscopy (AFM), CV, and electrochemical impedance spectroscopy (EIS). The obtained SEM and 3D-AFM images show that AuNPs deposited at the GC electrode prepared in NaNO3 solution form a significantly better, uniform, and homogeneous electrodeposited AuNP film on the GC electrode surface with nanoparticle sizes ranging from ∼36 to 60 nm. Meanwhile, from the electrochemical performances of the AuNP-modified GC electrodes, characterized by using a mixture of ferricyanide and ferrocyanide ions [Fe(CN6)3-/4-], there is no significant difference observed in the case of charge-transfer resistances (R ct) and heterogeneous electron-transfer rate constants (k o), although there are differences in the surface morphologies of the electrodeposited AuNP films. Remarkably, the R ct values of the AuNP-modified GC electrodes are lower than those of the bare GC electrode by 18-fold, as the R ct values were found to be ∼6 Ω (p < 0.001, n = 3). This has resulted in obtaining k o values of AuNP-modified GC electrodes between the magnitude of 10-2 and 10-3 cm s-1, giving a faster electron-transfer rate than that of the bare GC electrode (10-4 cm s-1). This study confirms that using an appropriate supporting background electrolyte plays a critical role in preparing electrodeposited AuNP films. This approach could lead to nanostructures with a more densely, uniformly, and homogeneously electrodeposited AuNP film on the electrode surfaces, albeit utilizing an easy and simple preparation method.

Dataset Information

A Nucleic Acid Nanostructure Built through On-Electrode Ligation for Electrochemical Detection of a Broad Range of Analytes.

Publications

A Nucleic Acid Nanostructure Built through On-Electrode Ligation for Electrochemical Detection of a Broad Range of Analytes.

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