This communication reports a nanocatalyst-based electrochemical assay for proteins. Ultrasensitive detection has been achieved by signal amplification combined with noise reduction: the signal is amplified both by the catalytic reduction of p-nitrophenol to p-aminophenol by gold-nanocatalyst labels and by the chemical reduction of p-quinone imine to p-aminophenol by NaBH4; the noise is reduced by employing an indium tin oxide electrode modified with a ferrocenyl-tethered dendrimer and a hydrophilic immunosensing layer.
The authors herein report optimized conditions for ultrasensitive phosphatase-based immunosensors (using redox cycling by a reducing agent) that can be simply prepared and readily applied to microfabricated electrodes. The optimized conditions were applied to the ultrasensitive detection of cardiac troponin I in human serum. The preparation of an immunosensing layer was based on passive adsorption of avidin (in carbonate buffer (pH 9.6)) onto indium-tin oxide (ITO) electrodes. The immunosensing layer allows very low levels of nonspecific binding of proteins. The optimum conditions for the enzymatic reaction were investigated in terms of the type of buffer solution, temperature, and concentration of MgCl(2), and the optimum conditions for antigen-antibody binding were determined in terms of incubation time, temperature, and concentration of phosphatase-conjugated IgG. Very importantly, the antigen-antibody binding at 4 °C is extremely important in obtaining reproducible results. Among the four phosphatase substrates (L-ascorbic acid 2-phosphate (AAP), 4-aminophenyl phosphate, 1-naphthyl phosphate, 4-amino-1-naphthyl phosphate) and four phosphatase products (L-ascorbic acid (AA), 4-aminophenol, 1-naphthol, 4-amino-1-naphthol), AAP and AA meet the requirements most for obtaining easy dissolution and high signal-to-background ratios. More importantly, fast AA electrooxidation at the ITO electrodes does not require modification with any electrocatalyst or electron mediator. Furthermore, tris(2-carboxyethyl)phosphine (TCEP) as a reducing agent allows fast redox cycling, along with very low anodic currents at the ITO electrodes. Under these optimized conditions, the detection limit of an immunosensor for troponin I obtained without redox cycling of AA by TCEP is ca. 100 fg/mL, and with redox cycling it is ca. 10 fg/mL. A detection limit of 10 fg/mL was also obtained even when an immunosensing layer was simply formed on a micropatterned ITO electrode. From a practical point of view, it is of great importance that ultralow detection limits can be obtained with simply prepared enzyme-based immunosensors.
We demonstrate a facile efficient way to fabricate activated carbon nanosheets (ACNSs) consisting of hierarchical porous carbon materials. Simply heating banana leaves with K2CO3 produce ACNSs having a unique combination of macro‐, meso‐ and micropores with a high specific surface area of ∼1459 m2 g−1. The effects of different electrolytes on the electrochemical supercapacitor performance and stability of the ACNSs are tested using a two‐electrode system. The specific capacitance (Csp) values are 55, 114, and 190 F g−1 in aqueous 0.5 M sodium sulfate, organic 1 M tetraethylammonium tetrafluoroborate in acetonitrile, and pure ionic liquid 1‐butyl‐3‐methylimidazolium hexafluorophosphate ([BMIM][PF6]) electrolytes, respectively. The ACNSs also shows the largest potential window of 3.0 V, the highest specific energy (59 Wh kg−1) and specific power (750 W kg−1) in [BMIM][PF6]. A mini‐prototype device is prepared to demonstrate the practicality of the ACNSs.
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