Paper Biosensor for the Detection of NT-proBNP Using Silver Nanodisks as Electrochemical Labels.

We report on the use of silver nanodisks (AgNDs), having a diameter of 50 ± 8 nm and a thickness of 8 ± 2 nm, as electrochemical labels for the detection of a model metalloimmunoassay for the heart failure biomarker NT-proBNP. The detection method is based on an electrochemically activated galvanic exchange (GE) followed by the detection of Ag using anodic stripping voltammetry (ASV). The AgNDs labels are superior to Ag nanocubes and Ag nanospheres in terms of the dynamic range for both the model and NT-proBNP metalloimmunoassays. The linear dynamic range for the model composite is 1.5 to 30.0 pM AgNDs. When AgND labels are used for the NT-proBNP assay, the dynamic range is 0.03-4.0 nM NT-proBNP. The latter range fully overlaps the risk stratification range for heart failure from 53 pM to 590 pM. The performance improvement of the AgNDs is a result of the specific GE mechanism for nanodisks. Specifically, GE is complete across the face of the AgNDs, leaving behind an incompletely exchanged ring structure composed of both Ag and Au.

Target-induced silver nanocluster generation for highly sensitive electrochemical aptasensor towards cell-secreted interferon-γ.

Interferon-gamma (IFN-γ) is one kind of crucial inflammatory cytokines, and its expression level is closely associated with various disease progressions. This work addresses the development of a sensitive and specific electrochemical assay for detection of IFN-γ by combing the recognition unit of aptamer with the signal reporter of target-induced silver nanoclusters (AgNCs). For biosensor preparation, the gold nanoparticles (AuNPs) immobilized on the amine-terminated electrode surface provided electrochemical interfaces for the self-assembly of C-rich modified aptamers. Then, the aptamer recognized IFN-γ and the free aptamer hybridized with conjugated DNA sequences. After the nuclease-catalyzed cleavage of DNA duplex, in situ-generated AgNCs in the C-rich template was utilized as the electrochemical indicator for IFN-γ detection. The present method demonstrated a good performance for detection of IFN-γ with a low detection limit of 1.7 pg mL-1. This aptasensor was verified to be applied for the evaluation of IFN-γ secreted by cell.

Effect of Serum on Electrochemical Detection of Bioassays Having Ag Nanoparticle Labels.

The effect of serum on electrochemical detection of bioassays having silver nanoparticle (AgNP) detection labels was investigated. Both a model assay and an antigen-specific sandwich bioassay for the heart failure marker NT-proBNP were examined. In both cases, the AgNP labels were conjugated to a detection antibody. Electrochemical detection was carried out using a galvanic exchange/anodic stripping voltammetry method in which Au3+ exchanges with AgNP labels. The assays were carried out using a paper-based electrode platform. The bioassays were exposed to different serum conditions prior to and during detection. There are three important outcomes reported in this article. First, both the model- and antigen-specific assays could be formed in undiluted serum with no detectable interferences from the serum components. Second, to achieve the maximum possible electrochemical signal, the highest percentage of serum that can remain in an assay buffer during electrochemical detection is 0.25% when no washing is performed. The assay results are rendered inaccurate when 0.50% or more of serum is present. Third, the factors inhibiting galvanic exchange in serum probably relate to surface adsorption of biomolecules onto the AgNP labels, chelation of Au3+ by serum components, or both. The results reported here provide general guidance for using metal NP labels for electrochemical assays in biofluids.

Silver Nanocubes as Electrochemical Labels for Bioassays.

Here, we report on the use of 40 ± 4 nm silver nanocubes (AgNCs) as electrochemical labels in bioassays. The model metalloimmunoassay combines galvanic exchange (GE) and anodic stripping voltammetry (ASV). The results show that a lower limit of detection is achieved by simply changing the shape of the Ag label yielding improved GE with AgNCs when compared to GE with spherical silver nanoparticles (sAgNPs). Specifically, during GE between electrogenerated Au3+ and the Ag labels, a thin shell of Au forms on the surface of the NP. This shell is more porous when GE proceeds on AgNCs compared to sAgNPs, and therefore, more exchange occurs when using AgNCs. ASV results show that the Ag collection efficiency (AgCE%) is increased by up to ∼57% when using AgNCs. When the electrochemical system is fully optimized, the limit of detection is 0.1 pM AgNCs, which is an order of magnitude lower than that of sAgNP labels.

Nitronyl nitroxide monoradical TEMPO as new electrochemical label for ultrasensitive detection of nucleic acids.

In this work, a novel electrochemical biosensor based on nitronyl nitroxide monoradical 2,2,6,6-tetramethylpiperidine 1-Oxyl (TEMPO) as new electrochemical label for facile nucleic acids detection is developed. This fast and convenient functional microelectrode was designed by fixing the capture probe peptide nucleic acid (PNA) and using the coordination interaction of Zr4+ with both phosphate groups and carboxyl groups. Differential pulse voltammetry (DPV) was used to study the oxidation current of TEMPO which was combined with the electrode surface and labeled. TEMPO electrochemical signal related to target deoxyribonucleic acid (tDNA) concentration was finally detected when tDNA was added on the surface of glassy carbon electrode (GCE). The detection principle, optimization of key factors and performance analysis of the biosensor are also discussed. A great linear relation is acquired within the scope of 10 pM-100 nM under optimal conditions and the detection limit of this experiment is calculated as low as 2.57 pM (R2 = 0.996). In addition, complex serum samples were used to explore the practical application of this experiment. The results show the developed electrochemical DNA biosensor has wide application prospects in nucleic acids detection and clinical analysis.

Label-free electrochemical immunosensor for insulin detection by high-efficiency synergy strategy of Pd NPs@3D MoSx towards H2O2.

Here, a novel H2O2-based electrochemical immunosensor utilizing Pd nanoparticles functionalized three-dimensional wrinkly amorphous MoSx composites (Pd NPs@3D MoSx) as the platform was developed for the determination of insulin. In this work, Pd NPs@3D MoSx prepared in the presence of CTAB possessed an excellent catalytic activity for the reduction of H2O2. Furthermore, Pd NPs@3D MoSx with favorable biological compatibility can conjugate a great many antibodies to capture insulin. Attributed to the excellent property, electrochemical signals could be greatly amplified, contributing to improving detection sensitivity. Especially, SEM, TEM, and XPS information further confirmed nanomaterial's surface morphology and amorphous structure. Under the optimal conditions, the proposed immunosensor exhibited a sensitively linear relation with logarithmic insulin concentrations from 0.01 to 100 ng/mL with a low detection limit of 3.0 pg/mL (S/N = 3). Characterized by good reproducibility, specificity, and stability, the fabricated immunosensor may blaze a path for insulin detection in a real sample.

A silver(I) doped bud-like DNA nanostructure as a dual-functional nanolabel for voltammetric discrimination of methylated from unmethylated genes.

A small DNA structure, referred to as DNA nanobud (NB), was used for the first time to design a dual-functional nanolabel in order to recognize a particular oligonucleotide sequence, generate and amplify the electrochemical analytical signal. NBs containing numerous repetitive desired sequences were prepared through self-assembly of 8-h rolling circle amplification. Then, redox-active silver ions were loaded onto the NBs by over-night incubation with a solution of AgNO3. The incorporation of Ag+ into NBs was confirmed by field emission scanning electron microscopy, dynamic light scattering, UV-Vis spectroscopy, zeta potential measurements, and energy-dispersive X-ray spectroscopy. A DNA sandwich complex was created after hybridization of Ag+-NB with target sequence, which was captured by immobilized probe on a gold electrode. Cyclic voltammetry was applied to measure the redox signal of silver ions produced typically at a potential around 0.02 V vs. Ag/AgCl. The label can specifically discriminate fully methylated BMP3 gene from fully unmethylated bisulfate-converted part of the gene. The electrochemical signal produced by DNA sandwich complex of gold/probe/BMP3/Ag+-NB was linear toward BMP3 concentration from 100 pM to 100 nM. The method has a 100 pM BMP3 detection limit. Conceivably, this nanolabel can be designed and modified such that it may also be used to detect other sequences with lower detection limits. Graphical abstract Ag+-NB as a new nanolabel for genosensing was formed by loading Ag+ on a spherical DNA nanostructure, nanobud (NB), synthesized by rolling circle amplification process. By using a gold electrode (AuE), Ag+-NB with numerous electroactive cations and binding sites can detect targets and generate amplified electrochemical signals.