Reduced graphene oxide (RGO) due to its excellent electrochemical properties and large surface area, has recently aroused much interest for electrochemical biosensing application. Here, the chemically active RGO has been synthesized and deposited onto an indium tin oxide (ITO) coated glass substrate by the electrophoretic deposition technique. This novel platform has been utilized for covalent attachment of the monoclonal antibodies of aflatoxin B1 (anti-AFB1) for food toxin (AFB1) detection. The electron microscopy, X-ray diffraction, and UV-visible studies reveal successful synthesis of reduced graphene oxide while the XPS and FTIR studies suggest its carboxylic functionalized nature. The electrochemical sensing results of the anti-AFB1/RGO/ITO based immunoelectrode obtained as a function of aflatoxin concentration show high sensitivity (68 μA ng(-1) mL cm(-2)) and improved detection limit (0.12 ng mL(-1)). The association constant (ka) for antigen-antibody interaction obtained as 5 × 10(-4) ng mL(-1) indicates high affinity of antibodies toward the antigen (AFB1).
There is a growing demand to integrate biosensors with microfluidics to provide miniaturized platforms with many favorable properties, such as reduced sample volume, decreased processing time, low cost analysis and low reagent consumption. These microfluidics-integrated biosensors would also have numerous advantages such as laminar flow, minimal handling of hazardous materials, multiple sample detection in parallel, portability and versatility in design. Microfluidics involves the science and technology of manipulation of fluids at the micro- to nano-liter level. It is predicted that combining biosensors with microfluidic chips will yield enhanced analytical capability, and widen the possibilities for applications in clinical diagnostics. The recent developments in microfluidics have helped researchers working in industries and educational institutes to adopt some of these platforms for point-of-care (POC) diagnostics. This review focuses on the latest advancements in the fields of microfluidic biosensing technologies, and on the challenges and possible solutions for translation of this technology for POC diagnostic applications. We also discuss the fabrication techniques required for developing microfluidic-integrated biosensors, recently reported biomarkers, and the prospects of POC diagnostics in the medical industry.
We report on a label-free microfluidic immunosensor with femtomolar sensitivity and high selectivity for early detection of epidermal growth factor receptor 2 (EGFR2 or ErbB2) proteins. This sensor utilizes a uniquely structured immunoelectrode made of porous hierarchical graphene foam (GF) modified with electrospun carbon-doped titanium dioxide nanofibers (nTiO2) as an electrochemical working electrode. Due to excellent biocompatibility, intrinsic surface defects, high reaction kinetics, and good stability for proteins, anatase nTiO2 are ideal for electrochemical sensor applications. The three-dimensional and porous features of GF allow nTiO2 to penetrate and attach to the surface of the GF by physical adsorption. Combining GF with functional nTiO2 yields high charge transfer resistance, large surface area, and porous access to the sensing surface by the analyte, resulting in new possibilities for the development of electrochemical immunosensors. Here, the enabling of EDC-NHS chemistry covalently immobilized the antibody of ErbB2 (anti-ErbB2) on the GF-nTiO2 composite. To obtain a compact sensor architecture, the composite working electrode was designed to hang above the gold counter electrode in a microfluidic channel. The sensor underwent differential pulse voltammetry and electrochemical impedance spectroscopy to quantify breast cancer biomarkers. The two methods had high sensitivities of 0.585 μA μM(-1) cm(-2) and 43.7 kΩ μM(-1) cm(-2) in a wide concentration range of target ErbB2 antigen from 1 × 10(-15) M (1.0 fM) to 0.1 × 10(-6) M (0.1 μM) and from 1 × 10(-13) M (0.1 pM) to 0.1 × 10(-6) M (0.1 μM), respectively. Utilization of the specific recognition element, i.e., anti-ErbB2, results in high specificity, even in the presence of identical members of the EGFR family of receptor tyrosine kinases, such as ErbB3 and ErbB4. Many promising applications in the field of electrochemical detection of chemical and biological species will derive from the integration of the porous GF-nTiO2 composite into microfluidic devices.
ABSTRACT:The surface modified and aligned mesoporous anatase titania nanofiber mats (TiO 2 −NF) have been fabricated by electrospinning for esterified cholesterol detection by electrochemical technique. The electrospinning and porosity of mesoporous TiO 2 −NF were controlled by use of polyvinylpyrrolidone (PVP) as a sacrificial carrier polymer in the titanium isopropoxide precursor. The mesoporous TiO 2 − NF of diameters ranging from 30 to 60 nm were obtained by calcination at 470°C and partially aligned on a rotating drum collector. The functional groups such as −COOH, −CHO etc. were introduced on TiO 2 −NF surface via oxygen plasma treatment making the surface hydrophilic. Cholesterol esterase (ChEt) and cholesterol oxidase (ChOx) were covalently immobilized on the plasma treated surface of NF (cTiO 2 −NF) via N-ethyl-N0-(3-dimethylaminopropyl carbodiimide) and N-hydroxysuccinimide (EDC-NHS) chemistry. The high mesoporosity (∼61%) of the fibrous film allowed enhanced loading of the enzyme molecules in the TiO 2 −NF mat. The ChEt-ChOx/cTiO 2 −NF-based bioelectrode was used to detect esterified cholesterol using electrochemical technique. The high aspect ratio, surface area of aligned TiO 2 −NF showed excellent voltammetric and catalytic response resulting in improved detection limit (0.49 mM). The results of response studies of this biosensor show excellent sensitivity (181.6 μA/mg dL −1 /cm 2 ) and rapid detection (20 s). This proposed strategy of biomolecule detection is thus a promising platform for the development of miniaturized device for biosensing applications.
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We report the fabrication of an efficient, label-free, selective and highly reproducible immunosensor with unprecedented sensitivity (femto-molar) to detect a breast cancer biomarker for early diagnostics. Mesoporous zinc oxide nanofibers (ZnOnFs) are synthesized by electrospinning technique with a fiber diameter in the range of 50-150 nm. Fragments of ZnOnFs are electrophoretically deposited on an indium tin oxide glass substrate and conjugated via covalent or electrostatic interactions with a biomarker (antiErbB2; epidermal growth factor receptor 2). Oxygen plasma treatment of the carbon doped ZnOnFs generates functional groups (-COOH, -OH, etc.) that are effective for the conjugation of anti-ErbB2. ZnOnFs without plasma treatment that conjugate via electrostatic interactions were also tested for comparison. Label-free detection of the breast cancer biomarker by this point-of-care device is achieved by an electrochemical impedance technique that has high sensitivity (7.76 kΩ µM ). This sensor is about an order of magnitude more sensitive than the best demonstrated in the literature based on different nanomaterials and about three orders of magnitude better than the ELISA standard for breast cancer biomarker detection. This proposed point-of-care cancer diagnostic offers several advantages, such as higher stability, rapid monitoring, simplicity, cost-effectiveness, etc., and should prove to be useful for the detection of other bio-and cancer markers.
There is an unmet need for improved fertilizer management in agriculture. Continuous monitoring of soil nitrate would address this need. This paper reports an all-solid-state miniature potentiometric soil sensor that works in direct contact with soils to monitor nitrate-nitrogen (NO3-N) in soil solution with parts-permillion (ppm) resolution. A working electrode is formed from a novel nanocomposite of poly(3-octylthiophene) and molybdenum disulfide (POT-MoS2) coated on a patterned Au electrode and covered with a nitrate-selective membrane using a robotic dispenser. The POT-MoS2 layer acts as an ion-to-electron transducing layer with high hydrophobicity and redox properties. The modification of the POT chain with MoS2 increases both conductivity and anion exchange, while minimizing the formation of a thin water layer at the interface between the Au electrode and the ion-selective membrane, which is notorious for solid-state potentiometric ion sensors. Therefore, the use of POT-MoS2 results in an improved sensitivity and selectivity of the working electrode. The reference electrode comprises a screen-printed silver/silver chloride (Ag/AgCl) electrode covered by a protonated Nafion layer to prevent chloride (Cl-) leaching in long-term measurements. This sensor was calibrated using both standard and extracted soil solutions, exhibiting a dynamic range that includes all concentrations relevant for agricultural applications (1-1500 ppm NO3-N). With the POT-MoS2 nanocomposite, the sensor offers a sensitivity of 64 mV/decade for nitrate detection, compared to 48 and 38 mV/decade for POT and MoS2 alone, respectively. The sensor was embedded into soil slurries where it accurately monitored nitrate for a duration of 27 days.
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