Nanostructured Cyclodextrin-Mediated Surface for Capacitive Determination of Cortisol in Multiple Biofluids.

The aim of this work is to develop a reusable polypropylene glycol (PPG):ß-cyclodextrin (ßCD) biosensor for cortisol detection. To achieve the most stable support for ßCD, we developed two PPG surfaces. The first surface is based on a gold surface modified with SAM of 3-mercaptopropionic acid (3MPA), and the second surface is based on a glassy carbon surface grafted with 4-carboxyphenyl diazonium salt. We characterized both surfaces by EIS, XPS, and ATR-FTIR and evaluated the stability and reusability of each surface. We found the GC-carboxyphenyl-PPG:ßCD is stable for at least 1 month. We have also demonstrated the reusability of the surface up to 10 times. In detecting cortisol, we used a nonfaradaic electrochemical impedance capacitive model to interpret the surface confirmation changes. We achieved sensitive detection of cortisol in PBS buffer, urine, and saliva with limit of detection of 2.13, 1.29, and 1.33 nM, respectively.

Perspective on Nanofiber Electrochemical Sensors: Design of Relative Selectivity Experiments.

The use of nanofibers creates the ability for non-enzymatic sensing in various applications and greatly improves the sensitivity, speed, and accuracy of electrochemical sensors for a wide variety of analytes. The high surface area to volume ratio of the fibers as well as their high porosity, even when compared to other common nanostructures, allows for enhanced electrocatalytic, adsorptive, and analyte-specific recognition mechanisms. Nanofibers have the potential to rival and replace materials used in electrochemical sensing. As more types of nanofibers are developed and tested for new applications, more consistent and refined selectivity experiments are needed. We applied this idea in a review of interferant control experiments and real sample analyses. The goal of this review is to provide guidelines for acceptable nanofiber sensor selectivity experiments with considerations for electrocatalytic, adsorptive, and analyte-specific recognition mechanisms. The intended presented review and guidelines will be of particular use to junior researchers designing their first control experiments, but could be used as a reference for anyone designing selectivity experiments for non-enzymatic sensors including nanofibers. We indicate the importance of testing both interferants in complex media and mechanistic interferants in the selectivity analysis of newly developed nanofiber sensor surfaces.

Application of Voltage in Dynamic Light Scattering Particle Size Analysis.

Dynamic light scattering (DLS) is a common method for characterizing the size distribution of polymers, proteins, and other nano- and microparticles. Modern instrumentation permits measurement of particle size as a function of time and/or temperature, but currently there is no simple method for performing DLS particle size distribution measurements in the presence of applied voltage. The ability to perform such measurements would be useful in the development of electroactive, stimuli-responsive polymers for applications such as sensing, soft robotics, and energy storage. Here, a technique using applied voltage coupled with DLS and a temperature ramp to observe changes in aggregation and particle size in thermoresponsive polymers with and without electroactive monomers is presented. The changes in aggregation behavior observed in these experiments were only possible through the combined application of voltage and temperature control. To obtain these results, a potentiostat was connected to a modified cuvette in order to apply voltage to a solution. Changes in polymer particle size were monitored using DLS in the presence of constant voltage. Simultaneously, current data were produced, which could be compared with particle size data, to understand the relationship between current and particle behavior. The polymer poly(N-isopropylacrylamide) (pNIPAM) served as a test polymer for this technique, as pNIPAM's response to temperature is well-studied. Changes in the lower-critical solution temperature (LCST) aggregation behavior of pNIPAM and poly(N-isopropylacrylamide)-block-poly(ferrocenylmethyl methacrylate), an electrochemically active block-copolymer, in the presence of applied voltage are observed. Understanding the mechanisms behind such changes will be important when trying to achieve reversible polymer structures in the presence of applied voltage.

Elucidating the Electrochemical Mechanism of NG-Hydroxy-L-arginine.

NG-Hydroxy-L-arginine (NOHA) is a stable intermediate product in the urea cycle that can be used to monitor the consumption of L-arginine by nitrous oxide synthase (NOS) to produce nitric oxide (NO) and L-citrulline. Research has implicated the urea cycle in many diseases and NO has cultivated interest as a potential biomarker for neural health. Electrochemical detection is an established, cost-effective method that can successfully detect low levels of analyte concentrations. As one of the few electrochemically active species in the urea cycle, NOHA shows promise as a biomarker for monitoring disruptions in this biochemical process. In this study, we show that NOHA has an oxidation peak at +355 mV vs Ag/AgCl at a glassy carbon electrode. In addition, cyclic voltammetry studies with structural analogs - alanine and N-hydroxyguanidine - allowed us to approximate the oxidation wave at +355 mV vs Ag/AgCl to be a one electron process. Diffusivity of NOHA was found using linear scan voltammetry with a rotating disk electrode and approximated at 5.50×10-5 cm2/s. Ample work is still needed to make a robust biosensor, but the results here characterize the electrochemical activity and represent principle steps in making a NOHA biosensor.

Reusable Cyclodextrin-Based Electrochemical Platform for Detection of trans-Resveratrol.

A reusable sensor architecture, through the combination of self-assembled monolayers and cyclodextrin supramolecular interactions, is demonstrated for class recognition of hydrophobic analytes demonstrated with trans-resveratrol. The reloadable sensor is based on reversible immobilization of α-cyclodextrin on polyethylene glycol surface. α-cyclodextrins complexes with polyethylene glycols and causes the polymer chains to change their surface configuration. The reproducibility and stability of the sur-face, in the detection of nanomolar concentrations of trans-resveratrol, can be demonstrated by electrochemical impedance spectroscopy, X-ray photoelectron spectroscopy, and Attenuated total reflectance-Fourier transform infrared spectroscopy. We propose that during sensor operation, α-cyclodextrin decouples from the poly-ethylene glycol surface to complex with trans-resveratrol in solution, and after use, the surface regeneration is conducted with a simple α-cyclodextrin soak. To test the nonspecific response, the sensor was also tested with trans-resveratrol spiked human urine.

Electrochemical characterization of the stimuli-response of surface-immobilized elastin-like polymers.

Elastin-like polymers (ELPs) are frequently used in a variety of bioengineering applications because of their stimuli-responsive properties. Above their transition temperature, ELPs will adopt different structures that promote intra- and intermolecular hydrophobic contacts to minimize unfavorable interactions with an aqueous environment. We electrochemically characterize the stimuli-responsive behavior of surface-immobilized ELPs corresponding to two proposed states: extended and collapsed. In the extended state the ELPs are more solvated. In the collapsed state, triggered by introducing an environmental stimulus, non-polar intramolecular contacts within ELPs are favored, resulting in quantifiable morphological changes on the surface characterized using electrochemical impedance spectroscopy (EIS). Charge transfer resistance, a component of impedance, was shown to increase after exposing an ELP modified electrode to a high salt concentration environment (3.0 M NaCl). An increase in charge transfer resistance indicates an increase in the insulating layer on the electrode surface consistent with the proposed mechanism of collapse, as the ELPs have undergone morphological changes to hinder the kinetics of the redox couple exchange. Further characterization of the surface-immobilized ELPs showed a reproducible surface modification, as well as reversibility and tunability of the stimuli-response.

Flow imaging microscopy as a novel tool for high-throughput evaluation of elastin-like polymer coacervates.

Biological and bioinspired polymer microparticles have broad biomedical and industrial applications, including drug delivery, tissue engineering, surface modification, environmental remediation, imaging, and sensing. Full realization of the potential of biopolymer microparticles will require methods for rigorous characterization of particle sizes, morphologies, and dynamics, so that researchers may correlate particle characteristics with synthesis methods and desired functions. Toward this end, we evaluated biopolymer microparticles using flow imaging microscopy. This technology is widely used in the biopharmaceutical industry but is not yet well-known among the materials community. Our polymer, a genetically engineered elastin-like polypeptide (ELP), self-assembles into micron-scale coacervates. We performed flow imaging of ELP coacervates using two different instruments, one with a lower size limit of approximately 2 microns, the other with a lower size limit of approximately 300 nanometers. We validated flow imaging results by comparison with dynamic light scattering and atomic force microscopy analyses. We explored the effects of various solvent conditions on ELP coacervate size, morphology, and behavior, such as the dispersion of single particles versus aggregates. We found that flow imaging is a superior tool for rapid and thorough particle analysis of ELP coacervates in solution. We anticipate that researchers studying many types of microscale protein or polymer assemblies will be interested in flow imaging as a tool for quantitative, solution-based characterization.

Evaluation of Metal Oxide Surface Catalysts for the Electrochemical Activation of Amino Acids.

Electrochemical detection of amino acids is important due to their correlation with certain diseases; however, most amino acids require a catalyst to electrochemically activate. One common catalyst for electrochemical detection of amino acids are metal oxides. Metal oxide nanoparticles were electrodeposited onto glassy carbon and platinum working electrodes. Cyclic voltammetry (CV) experiments in a flow cell were performed to evaluate the sensors' ability to detect arginine, alanine, serine, and valine at micromolar and nanomolar concentrations as high as 4 mM. Solutions were prepared in phosphate buffer saline (PBS) and then 100 mM NaOH. Specifically, NiO surfaces were responsive to amino acids but variable, especially when exposed to arginine. Polarization resistance experiments and scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) data indicated that arginine accelerated the corrosion of the NiO catalyst through the formation of a Schiff base complex.

Guide to Selecting a Biorecognition Element for Biosensors.

Biosensors are powerful diagnostic tools defined as having a biorecognition element for analyte specificity and a transducer for a quantifiable signal. There are a variety of different biorecognition elements, each with unique characteristics. Understanding the advantages and disadvantages of each biorecognition element and their influence on overall biosensor performance is crucial in the planning stages to promote the success of novel biosensor development. Therefore, this review will focus on selecting the optimal biorecognition element in the preliminary design phase for novel biosensors. Included is a review of the typical characteristics and binding mechanisms of various biorecognition elements, and how they relate to biosensor performance characteristics, specifically sensitivity, selectivity, reproducibility, and reusability. The goal is to point toward language needed to improve the design and development of biosensors toward clinical success.

Controlling the Sensing Properties of Silicon Nanowires via the Bonds Nearest to the Silicon Nanowire Surface.

Controlling the sensing properties of a silicon nanowire field effect transistor is dependent on the surface chemistry of the silicon nanowire. A standard silicon nanowire has a passive oxide layer (native oxide), which has trap states that cause sensing inaccuracies and desensitize the surface to nonpolar molecules. In this paper, we successfully modified the silicon nanowire surface with different nonoxide C3 alkyl groups, specifically, propyl (Si-CH2-CH2-CH3), propenyl (Si-CH═CH-CH3), and propynyl (Si-C≡C-CH3) modifications. The effect of the near surface bond on the sensor sensitivity and stability was explored by comparing three C3 surface modifications. A reduction of trap-states led to greater sensor stability and accuracy. The propenyl-modified sensor was consistently the most stable and sensitive sensor, among the applied sensors. The propenyl- and propynyl-modified sensors consistently performed with the best accuracy in identifying specific analytes with similar polarity or similar molecular weights. A combination of features from different sensing surfaces led to the best rubric for specific analytes identification. These results indicate that nonoxide sensor surfaces are useful in identifying specific analytes and that a combination of sensors with different surfaces in a cross-reactive array can lead to specific analytes detection.