RESUMEN
Solid-state nanopore has the ability to detect proteins at a single-molecule level with its high sensitivity, high-throughput, and low cost. Improvements in fabrication, functionalization, and characterization of solid-state nanopores keep evolving. Various analytical methods targeted towards diagnostic applications using nanopore-based devices are appearing. This review article provides an overview of recent progress in the field of solid-state and biological nanopores for protein detection in a complex analyte. The advantages and challenges involved in nanopore sensing have been discussed. Further, the review surpasses the steady-state resistive pulse techniques of sensing and incorporate transient variations in the nanopore conductance. Application of the power spectral density of these fluctuations toward sensing has been highlighted with importance on reducing the detection limit in a complex environment. Lastly, the current problems and future perspectives have been discussed with a perspective to increase nanopores performance towards diagnostic applications in complex medium.
Asunto(s)
Nanoporos , Nanotecnología , ProteínasRESUMEN
To enable detection and discovery of biomarkers, development of label-free, ultrasensitive, and specific sensors is the need of the hour. For addressing this requirement, here, a Schottky-contacted ZnO nanorod biosensor has been demonstrated, which explores the interplay between Schottky junction capacitance and solution resistance, resulting in an interesting sensing principle of competitive impedance spectroscopy. When the transition of dominating impedance occurs from solution resistance to junction capacitance, a notch or a peak appears in the impedance response at a particular frequency (referred to as the corner frequency) depending on the charge of the target molecule. The appearance of the peak or notch acts like an electronic label for selectivity since it is visible only for target molecules even at ultralow concentrations in the physiological analyte, where the magnitude of impedance change overlaps with that for nonspecific molecules. This phenomenon has been successfully applied for the positively charged vascular endothelial growth factor (VEGF) and the negatively charged hepatitis B surface antigen (HBsAg), where the shifts in the higher corner frequencies for 1 aM concentration of the target molecules have been observed to be more than 3 times the changes in the impedance magnitude. Further, the area of the ZnO nanorods was segmented into two zones corresponding to the lower and higher concentration regimes, thereby expanding the dynamic range. To summarize, an ultralow detection limit of 1 aM with a dynamic range up to 1 pM was achieved for VEGF and HBsAg, which is 4 orders of magnitude and 20 times lower than their most sensitive label-free reports, respectively.
Asunto(s)
Nanotubos , Óxido de Zinc , Espectroscopía Dieléctrica , Antígenos de Superficie de la Hepatitis B , Nanotubos/química , Factor A de Crecimiento Endotelial Vascular , Óxido de Zinc/químicaRESUMEN
Affordable point-of-care (PoC) diagnostic devices enable detection of prostate specific antigen (PSA) in resource limited settings. Despite the advancements in PoC systems, most of the reported methods for PSA detection have unsatisfactory detection limits and are based on labelled assays, requiring multiple reagent flow steps which increases both expenses and inconvenience. Circumventing these constraints, we report here the development and validation of a label free, affordable dielectrophoresis (DEP) based graphene field effect transistor (FET) sensor implemented using coplanar electrodes and integrated uniquely with a compact disc based microfluidic platform along with electronics readout for the estimation of PSA at the point of care. Design of coplanar gate electrode which has not been explored earlier is not a straightforward approach. In fact, it has been observed that there is a non-monotonic dependence of the capture of PSA molecules in the channel region of the FET with varying widths and spacings of the gate electrode. The graphene FET based PoC device with optimized coplanar gate electrode is the only label free analytical system for PSA detection requiring simple operation and achieving a detection limit of 1 pg/ml in serum with a wide dynamic range upto 4 ng/ml and appreciable selectivity against potential interferents like bovine serum albumin (BSA) and human immunoglobulin G (IgG). Further, it has been validated satisfactorily with commercially available existing systems using human serum samples. Moreover, the proposed sensing system lowers the detection limit by three orders of magnitude compared to a recent study on label free PoC device on other cancer biomarkers.
Asunto(s)
Técnicas Biosensibles , Antígeno Prostático Específico/análisis , Electrodos , Grafito , Humanos , Masculino , Microfluídica , Sistemas de Atención de Punto , Pruebas en el Punto de AtenciónRESUMEN
An understanding of the current change in a solid state nanopore due to particle movement or capture is crucial for improvement of nanopore based sensing technologies. For lower aspect ratio pores, which are gaining importance due to their high sensitivity, there is interplay between access and pore resistance and the existing theories for computation of access resistance cannot explain most of the experimental observations. Hence, there is a need to develop a comprehensive model for calculating the effective conductance drop in presence of particles in a solid state nanopore. In this paper, we develop analytical models to calculate both the access and pore resistance in presence of particle at different positions during translocation and also when captured by receptors in functionalized nanopores. A wide range of pore geometry and molar strength has been investigated. Taking into consideration the positional uncertainty during particle translocation, the effective resistance sensitivity has been found to agree very well with the experimental observations in low aspect ratio pore. Additionally, we observe that in functionalized nanopores, a pore of higher diameter results in around 50% increase in sensitivity compared to a pore with half its diameter, which indicates the scope of design optimization in such systems.