Nanostructured titanium nitride for highly sensitive localized surface plasmon resonance biosensing

Titanium nitride (TiN) as an alternative plasmonic ceramic material with superb properties including high hardness, outstanding corrosion resistance and excellent biocompatibility, has exhibited great potential for optical biochemical sensing applications. By sputtering about 35 nm–50 nm TiN on glass (f-TiN), the surface was found to provide sensing capability toward NaCl solution through the phenomenon of surface plasmon resonance. When the TiN film of about 27 nm–50 nm in thickness was sputtered onto a roughened glass surface (R–TiN), the sensing capability was improved. This was further improved when holes at nanoscale were created in the TiN film of about 19 nm–27 nm in thickness (NH–TiN). The roughened surface and nanohole patterns provided confinement of surface plasmons and significantly improved the sensitivity toward the local refractive index changes. In detail, the calculated refractive index resolution (RIR) of the optimal NH–TiN sensors for NaCl was found to be 9.5 × 10⁻⁸ refractive index unit (RIU), which had outperformed the f-TiN and R–TiN sensors. For biosensing, the optimized NH–TiN sensor was found to be capable to detect both small and large biomolecules, i.e. biotin (molecular weight of 244.3 g/mol) and human IgG (160,000 g/mol), in a label-free manner. Especially, the NH–TiN sensor significantly improved sensitivity in detecting small molecules due to the localized plasmonic confinement of electromagnetic field. Combining with the excellent mechanical and durability properties of TiN, the proposed NH–TiN can be a strong candidate for plasmonic biosensing applications.     

Interfacial hybridization kinetics of oligonucleotides immobilized onto fused silica surfaces

Fused silica optical fibers have been used in an intrinsic mode optical configuration as biosensors for fluorescence based detection of hybridization of nucleic acids. In this work, the kinetics of hybridization of single-stranded oligonucleotides that were covalently immobilized were studied. The probe DNA was dT₂₀, and the target was Fluorescein-labeled non-complementary (dT₂₀) or complementary (dA₂₀) oligonucleotide. Chronofluorimetric monitoring of the adsorption and hybridization processes was used to investigate oligonucleotide films of different density, in different salt concentrations, at temperatures of 25 and 40 °C, with the concentration of the target DNA being 0.005–0.1 μM. Mathematical models based on first- and second-order Langmuir adsorption have been examined to describe both the adsorption and the hybridization processes. Experimental data were processed using the models, and the hybridization kinetics were calculated. Hybridization kinetics on these optical fiber DNA sensors was found to be up to three orders faster than results presented for a number of other experiments using different immobilization chemistries.     

A new amperometric biosensor for fructose determination based on epoxy-graphite-TTF-TCNQ-FDH-biocomposite

In this work a novel amperometric biosensor for fructose determination in solutions was developed. The device was constructed by the incorporation of a tetrathiofulvalene-tetracyanoquinodimethane organic conducting salt and fructose dehydrogenase enzyme, include in a polymeric matrix of epoxy resin and graphite powder. Because of the electrocatalytic function of the salt, the direct transfer of the electron between the reduced prosthetic group (PQQH₂) of the enzyme and the transducing material, was verified at a low working potential (150 mV vs. Ag/AgCl), where the interfering reactions were minimized. The response time at 90% of the steady state value was less than 20 s. The current response was directly proportional to the D-fructose concentration from 0.01 to 0.3 mmol/l with a detection limit of 0.005 mmol/l (signal/noise of 3) and a sensitivity of 1.9985 μA/mmol. The biosensor sensitivity diminishes when its surface is not polished between successive determinations, and remains constant (rsd=1.85, n=10) when the surface is polished between determinations. The effects of temperature and pH on the biosensor response were studied and analyzed; also the properties of the enzyme (Km _ap, I _max, Q₁₀) were determinate in this work. The biosensor was used to determine fructose in high fructose syrups and there were not significant differences between these results and those obtained by HPLC (p≤0.05). During 4 months, in intermittent determinations the biosensor kept 100% of its original sensitivity and after 18 months stored at 4°C, it only lost 32% of its sensitivity. The simplicity, low working potential, high stability and good performance of this biosensor shows a great potential for its use in the fructose determination.     

Generation of a green fluorescent protein gene chromosomal insertion containing Escherichia coli strain for gene induction-based quantification of bioavailable lysine

The importance of lysine determination in feed materials is crucial for the feed industry because this amino acid can be limiting in many of the cereal materials used for animal feeds. The bacterial gene induction-based assay developed in this study aimed to measure lysine bioavailability in feeds as an alternative analytical method for animal assays. The advantages of a gene induction-based approach include rapid and quantitative estimation of many samples and results that relate a bacterial response to a biological response observed in animals. A whole-cell biosensor strain was constructed using a fluorescent E. coli strain that has an inducible fluorescent phenotype sensitive to extracellular lysine contents. A genetic fusion that links the promoter of cad operon with a green fluorescent protein encoding gene (gfp) was constructed, and a fluorescent assay was developed. A standard lysine curve (R² = 0.95) was generated and used for lysine bioavailability quantification of four feed ingredients (whole egg protein, blood-, soybean-, and meat and bone meal). Quantities as low as 50 μg/ml protein of digested samples were sufficient for analyses using the biosensor, except for meat and bone meal. Because of the low levels of free lysine in non-digested samples, fluorescence of these protein sources containing lower than 500 μg/ml protein was not detected (except for soybean meal). The results using enzymatically digested protein sources showed that the test strain emitted a fluorescent response that was proportional to the level of lysine present in the feed samples.     

A shear horizontal surface acoustic wave sensor for the detection of antigen–antibody reactions for medical diagnosis

In this paper we describe the development of a shear horizontal surface acoustic wave immunosensor for medical diagnostics. There exists a strong interest in the rapid detection of antigen–antibody reactions at small concentrations in the g/ml region in plasma, serum, or whole blood. Sensors whose operating principle is based on shear horizontal acoustic surface waves are well suited for this. We have used a spin-on glass film for the guidance of the surface wave as well as for the protection of the aluminum structures of the surface wave transducers from aggressive analyte liquids. This film has proven to considerably enhance the sensitivity of the device, and to simultaneously provide a durable protection of the transducers. Furthermore, polymers based on polyvinylamines have been used for the first time for immobilization of the capture protein. This technique effectively prevents the undesired binding of foreign substances like cells, non-specific antibodies, or other proteins at the sensor surface.     

Potentiometric urea biosensor based on BSA embedded surface modified polypyrrole film

A potentiometric biosensor based on bovine serum albumin (BSA) embedded surface modified polypyrrole has been developed for the quantitative estimation of urea in aqueous solution. The enzyme, urease (Urs), was covalently linked to free amino groups present over the BSA embedded modified surface of the conducting polypyrrole film electrochemically deposited onto an indium–tin-oxide (ITO) coated glass plate. The biosensor has been characterized by UV–visible, infrared spectroscopy and SEM. Potentiometric and spectrophotometric response of the enzyme electrode (Urs/BSA-PPy/ITO) were measured as a function of urea concentration in Tris–HCl buffer (pH 7.0). It has been found that the electrode responds to low urea concentration with wider range of detection. The electrode showed a linear response range of 6.6 × 10⁻⁶ to 7.5 × 10⁻⁴ M urea. The response time is about 70–90 s reaching to a 95% steady-state potential value and 75% of the enzyme activity is retained for about 2 months. These results indicate an efficient covalent linkage of enzyme to free amino groups of the BSA molecules over the surface of polypyrrole film, which leads to high enzyme loading, an increased lifetime stability of the electrode and an improved wide range of detection of low urea concentration in aqueous solution.     

Controlled chemical modification of the internal surface of photonic crystal fibers for application as biosensitive elements

Photonic crystal fibers (PCF) are one of the most promising materials for creation of constructive elements for bio-, drug and contaminant sensing based on unique optical properties of the PCF as effective nanosized optical signal collectors. In order to provide efficient and controllable binding of biomolecules, the internal surface of glass hollow core photonic crystal fibers (HC-PCF) has been chemically modified with silanol groups and functionalized with (3-aminopropyl) triethoxysilane (APTES). The shift of local maxima in the HC-PCF transmission spectrum has been selected as a signal for estimating the amount of silanol groups on the HC-PCF inner surface. The relationship between amount of silanol groups on the HC-PCF inner surface and efficiency of following APTES functionalization has been evaluated. Covalent binding of horseradish peroxidase (chosen as a model protein) on functionalized PCF inner surface has been performed successively, thus verifying the possibility of creating a biosensitive element.     

A high-density multi-point LAPS set-up using a VCSEL array and FPGA control

A new LAPS (light-addressable potentiometric sensor) set-up will be introduced, in which the light sources are miniaturised by the utilisation of a VCSEL (vertical-cavity surface-emitting laser) array to increase the measurement spot density. An FPGA (field-programmable gate array) is used to generate modulation signals for individual illumination of each measurement spot of the LAPS. The new set-up can operate a large number of measurement spots simultaneously by reading out the sum photocurrent and separate the signals of the individual measurement spots by an FFT analysis. The frequency, amplitude and offset of the modulation signal can be configured for each measurement spot by software. The new system can be combined with a positioning stage allowing the parallel read out of a single line of measurement spots and a scan perpendicular to that line in a similar manner, like for an optical scanner set-up. First measurements demonstrate the functionality of the new LAPS set-up as a chemical imaging system.     

Enhanced surface plasmon resonance detection of DNA hybridization based on ZnO nanorod arrays

We demonstrated an enhanced surface plasmon resonance detection incorporating ZnO nanorod arrays (NRAs) built on a thin gold film. ZnO NRAs were fabricated by wet chemical growth method and used for the detection of DNA hybridization. Experimental results exhibited that ZnO NRAs provided a notable sensitivity improvement by more than 3 times, which is attributed to an increase in the surface reaction area. The measured sensitivity enhancement matched well with the numerical analyses based on the effective medium theory. Our approach is intended to show the feasibility and extend the applicability of the ZnO-based SPR biosensor to diverse biomolecular binding events.     

Multiwalled carbon nanotubes grafted chitosan nanobiocomposite: A prosperous functional nanomaterials for glucose biosensor application

Multiwalled carbon nanotubes (MWNTs) grafted chitosan (CS) nanowire (NW) was prepared by phase separation method. Glucose oxidase (GOx) was sequentially immobilized into MWNT-CS-NW to obtain MWNT-CS-NW/GOx biosensor. Field emission scanning electron microscopy (FESEM) images of MWNT-CS-NW/GOx reveals the existence of MWNT and CS. Cyclic voltammetry and amperometry were used to evaluate the electrochemical determination of glucose. The MWNT-CS-NW/GOx biosensor shows an excellent performance for glucose at +0.34 V with a high sensitivity (5.03 μA/mM) and lower response time (3 s) in a wide concentration range of 1-10 mM (correlation coefficient of 0.9988). In addition, MWNT-CS-NW/GOx biosensor possesses better reproducibility, storage stability and there is negligible interference from other electroactive components.