Due to their all-electrical nature, impedance biosensors have significant potential for use as simple and portable sensors for environmental studies and environmental monitoring. Detection of two endocrine-disrupting chemicals (EDC), norfluoxetine and BDE-47, is reported here by impedance biosensing, with a detection limit of 8.5 and 1.3 ng/mL for norfluoxetine and BDE-47, respectively. Although impedance biosensors have been widely studied in the academic literature, commercial applications have been hindered by several technical limitations, including possible limitations to small analytes, the complexity of impedance detection, susceptibility to nonspecific adsorption, and stability of biomolecule immobilization. Recent research into methods to overcome these obstacles is briefly reviewed. New results demonstrating antibody regeneration atop degenerate (highly doped) Si are also reported. Using 0.2 M KSCN and 10 mM HF for antibody regeneration, peanut protein Ara h 1 is detected daily during a 30 day trial.
An antibody-based analytical method for the detection of a chemical flame retardant using antibody fragments isolated from an alpaca has been developed. One specific chemical flame retardant congener, 2,2′,4,4′-tetrabrominated diphenyl ether (BDE-47), is often the major poly-BDE (PBDE) congener present in human and environmental samples and that which is the most frequently detected. An alpaca was immunized with a surrogate of BDE-47 covalently attached to a carrier protein. The resulting mRNA coding for the variable domain of heavy-chain antibodies (VHH) were isolated, transcribed to cDNA, and cloned into a phagemid vector for phage display library construction. Selection of VHHs recognizing BDE-47 was achieved by panning under carefully modified conditions. The assay sensitivity for detecting BDE-47 was down to the part-per-billion (microgram per liter) level. Cross-reactivity analyses confirmed that this method was highly selective for BDE-47 and selected hydroxylated metabolites. When exposed to elevated temperatures, the camelid VHH antibodies retained more reactivity than a polyclonal antibody developed to the same target analyte. The use of this VHH antibody reagent immobilized onto a Au electrode for impedance biosensing demonstrates the increased versatility of VHH antibodies.
Food pathogens contaminate food products that allow their growth on the shelf and also under refrigerated conditions. Therefore, it is of utmost importance to lower the limit of detection (LOD) of the method used and to obtain the results within hours to few days. Biosensor methods exploit the available technologies to individuate and provide an approximate quantification of the bacteria present in a sample. The main bottleneck of these methods depends on the aspecific binding to the surfaces and on a change in sensitivity when bacteria are in a complex food matrix with respect to bacteria in a liquid food sample. In this review, we introduce surface plasmon resonance (SPR), new advancements in SPR techniques, and electrochemical impedance spectroscopy (EIS), as fluorescence-free biosensing technologies for detection of L. monocytogenes in foods. The application of the two methods has facilitated L. monocytogenes detection with LOD of 1 log CFU/mL. Further advancements are envisaged through the combination of biosensor methods with immunoseparation of bacteria from larger volumes, application of lab-on-chip technologies, and EIS sensing methods for multiplex pathogen detection. Validation efforts are being conducted to demonstrate the robustness of detection, reproducibility and variability in multi-site installations.
The thermal behavior of carbon-coated LiFePO4 cathode material was investigated using three different calorimetric techniques. Heat released during charge and discharge of cylindrical 18650 size cells was measured in an isothermal calorimeter. Accelerating rate calorimetry was used to investigate the cause of thermal runaway of the cylindrical 18650 size cells. On the other hand, differential scanning calorimeter was used to study the contribution of the delithiated carbon-coated LiFePO4 and the lithiated graphite to the overall thermal runaway through coin cells of 2032 size. The carbon-coated LiFePO4 showed very good thermal stability.
Mild denaturing agents were studied for antibody regeneration atop degenerate (highly doped) Si and Au electrodes using comparable 11-carbon chain linker reagents onto which the mouse monoclonal antibody to peanut protein Ara h 1 is covalently immobilized. Of the reagents studied, only 200mM KSCN is effective for antibody regeneration and detection of Ara h 1 atop Au electrodes, allowing 15days of sensor usage after daily antibody unfolding and refolding, while 200mM KSCN +10mM HF is effective atop degenerate Si electrodes, allowing 30days of sensor usage. The addition of HF is required for antibody regeneration atop Si electrodes, as demonstrated by cyclic voltammetry in the presence of 5.0mM K3Fe(CN)6/K4Fe(CN)6 at pH7.3, where the oxidation/reduction peaks can be observed only in the presence of HF. The impedance spectrum for detection of Ara h 1 gradually degrades during these multi-day regeneration trials, but calibration experiments performed within one day illustrate that sensor electrodes can be calibrated on the day of use to within about 2%
We report an impedance biosensor utilizing a Si electrode created by wet chemical deposition atop Al alloy 6061. The sensor electrode is created by galvanic/electroless Si deposition from an electrolyte containing 10 mM HF and 20 mM Na2SiF6 in 80 wt% formic acid, followed by antibody immobilization. The impedance response of the sensor electrode to increasing concentrations of peanut protein Ara h 1, a common food allergen, can be fit to an equivalent circuit containing three RC loops. The circuit element most sensitive to antigen binding is the charge transfer resistance, yielding a detection limit of 4 ng/ml.