Investigation of the Morphology and Electrical Properties of Graphene Used in the Development of Biosensors for Detection of Influenza Viruses

In this study, we discuss the mechanisms behind changes in the conductivity, low-frequency noise, and surface morphology of biosensor chips based on graphene films on SiC substrates during the main stages of the creation of biosensors for detecting influenza viruses. The formation of phenylamine groups and a change in graphene nano-arrangement during functionalization causes an increase in defectiveness and conductivity. Functionalization leads to the formation of large hexagonal honeycomb-like defects up to 500 nm, the concentration of which is affected by the number of bilayer or multilayer inclusions in graphene. The chips fabricated allowed us to detect the influenza viruses in a concentration range of 10−16 g/mL to 10−10 g/mL in PBS (phosphate buffered saline). Atomic force microscopy (AFM) and scanning electron microscopy (SEM) revealed that these defects are responsible for the inhomogeneous aggregation of antibodies and influenza viruses over the functionalized graphene surface. Non-uniform aggregation is responsible for a weak non-linear logarithmic dependence of the biosensor response versus the virus concentration in PBS. This feature of graphene nano-arrangement affects the reliability of detection of extremely low virus concentrations at the early stages of disease.

Modified Nanodiamonds as a Means of Polymer Surface Functionalization. From Fouling Suppression to Biosensor Design

The development of different methods for tuning surface properties is currently of great interest. The presented work is devoted to the use of modified nanodiamonds to control the wetting and biological fouling of polymers using optical sensors as an example. We have shown that, depending on the type of modification and the amount of nanodiamonds, the surface of the same fluorinated polymer can have both bactericidal properties and, on the contrary, good adhesion to the biomaterial. The precise control of wetting and biofouling properties of the surface was achieved by the optimization of the modified nanodiamonds thermal anchoring conditions. In vitro and in vivo tests have shown that the fixation of amine functional groups leads to inhibition of biological activity, while the presence of a large number of polar groups of mixed composition (amide and acid chloride) promotes adhesion of the biomaterial and allows one to create a biosensor on-site. A comprehensive study made it possible to establish that in the first 5 days the observed biosensor response is provided by cells adhered to the surface due to the cell wall interaction. On the 7th day, the cells are fixed by means of the polysaccharide matrix, which provides much better retention on the surface and a noticeably greater response to substrate injections. Nevertheless, it is important to note that even 1.5 h of incubation is sufficient for the formation of the reliable bioreceptor on the surface with the modified nanodiamonds. The approach demonstrated in this work makes it possible to easily and quickly isolate the microbiome on the surface of the sensor and perform the necessary studies of its substrate specificity or resistance to toxic effects.

Understanding the Mechanism of Formation of a Response to Juglone for Intact and Immobilized Bacterial Cells as Recognition Elements of Microbial Sensors: Processes Causing the Biosensor Response

Microbial reactor sensors (based on freshly harvested intact microbial cells) or microbial membrane sensors (based on immobilized microbial cells) can be used as convenient instruments for studying processes that cause the response of a biosensor, such as the properties of enzymes or the characteristics of metabolism. However, the mechanisms of the formation of biosensors responses have not yet been fully understood to study only one of these processes. In this work, the results of studies on the formation of a response to juglone for intact and immobilized bacterial cells used as receptors are presented. It was shown that the contribution of reactive oxygen species (ROS) to the formation of the biosensor response depends on the culture receptor and the form of juglone, quinone, or phenolate used. The response to the quinone form of juglone both for intact and immobilized cells of catalase-positive actinobacterium is formed regardless of the presence of ROS. The response of freshly harvested intact actinobacterial cells was caused by the rate of the enzymatic conversion of juglone. The rate of the response of immobilized actinobacterial cells was influenced by the activity of transport systems and metabolism. The response of immobilized pseudomonad cells was caused by the transport of juglone into cells, the inhibitory effect of juglone-induced ROS, and juglone metabolism.

Ultrasensitive RNA biosensors for SARS-CoV-2 detection in a simple color and luminescence assay

The COVID-19 pandemic underlines the need for versatile diagnostic strategies. Here, we have designed and developed toehold RNA-based sensors for direct and ultrasensitive SARS-CoV-2 RNA detection. In our assay, isothermal amplification of a fragment of SARS-CoV-2 RNA coupled with activation of our biosensors leads to a conformational switch in the sensor. This leads to translation of a reporter-protein e.g. LacZ or Nano-lantern that is easily detected using color/luminescence. This response can be visualized by the human eye, or a simple cell phone camera as well as quantified using a spectrophotometer/luminometer. By optimizing RNA-amplification and biosensor-design, we have generated a highly-sensitive diagnostic assay; with sensitivity down to attomolar (100 copies of) SARS-CoV-2 RNA. Finally, this PHAsed NASBA-Translation Optical Method (PHANTOM) efficiently detects the presence of viral RNA in human patient samples, with clear distinction from samples designated negative for the virus. The biosensor response correlates well with Ct values from RT-qPCR tests and thus presents a powerful and easily accessible strategy for detecting Covid infection.

Layered Double Hydroxide-Modified Organic Electrochemical Transistor for Glucose and Lactate Biosensing

Biosensors based on Organic Electrochemical Transistors (OECTs) are developed for the selective detection of glucose and lactate. The transistor architecture provides signal amplification (gain) with respect to the simple amperometric response. The biosensors are based on a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) channel and the gate electrode is functionalised with glucose oxidase (GOx) or lactate oxidase (LOx) enzymes, which are immobilised within a Ni/Al Layered Double Hydroxide (LDH) through a one-step electrodeposition procedure. The here-designed OECT architecture allows minimising the required amount of enzyme during electrodeposition. The output signal of the biosensor is the drain current (Id), which decreases as the analyte concentration increases. In the optimised conditions, the biosensor responds to glucose in the range of 0.1–8.0 mM with a limit of detection (LOD) of 0.02 mM. Two regimes of proportionality are observed. For concentrations lower than 1.0 mM, a linear response is obtained with a mean gain of 360, whereas for concentrations higher than 1.0 mM, Id is proportional to the logarithm of glucose concentration, with a gain of 220. For lactate detection, the biosensor response is linear in the whole concentration range (0.05–8.0 mM). A LOD of 0.04 mM is reached, with a net gain equal to 400.

An Optical Urate Biosensor Based on Urate Oxidase and Long-Lifetime Metalloporphyrins

Gout is a condition that affects over 8 million Americans. This condition is characterized by severe pain, and in more advanced cases, bone erosion and joint destruction. This study explores the fabrication and characterization of an optical, enzymatic urate biosensor for gout management, and the optimization of the biosensor response through the tuning of hydrogel matrix properties. Sensors were fabricated through the co-immobilization of oxygen-quenched phosphorescent probes with an oxidoreductase within a biocompatible copolymer hydrogel matrix. Characterization of the spectral properties and hydrogel swelling was conducted, as well as evaluation of the response sensitivity and long-term stability of the urate biosensor. The findings indicate that increased acrylamide concentration improved the biosensor response by yielding an increased sensitivity and reduced lower limit of detection. However, the repeatability and stability tests highlighted some possible areas of improvement, with a consistent response drift observed during repeatability testing and a reduction in response seen after long-term storage tests. Overall, this study demonstrates the potential of an on-demand, patient-friendly gout management tool, while paving the way for a future multi-analyte biosensor based on this sensing platform.

An Electrochemical DNA Biosensor for Carcinogenicity of Anticancer Compounds Based on Competition between Methylene Blue and Oligonucleotides

A toxicity electrochemical DNA biosensor has been constructed for the detection of carcinogens using 24 base guanine DNA rich single stranded DNA, and methylene blue (MB) as the electroactive indicator. This amine terminated ssDNA was immobilized onto silica nanospheres and deposited on gold nanoparticle modified carbon-paste screen printed electrodes (SPEs). The modified SPE was initially exposed to a carcinogen, followed by immersion in methylene blue for an optimized duration. The biosensor response was measured using differential pulse voltammetry. The performance of the biosensor was identified on several anti-cancer compounds. The toxicity DNA biosensor demonstrated a linear response range to the cadmium chloride from 0.0005 ppm to 0.01 ppm (R2 = 0.928) with a limit of detection at 0.0004 ppm. The biosensor also exhibited its versatility to screen the carcinogenicity of potential anti-cancer compounds.

Construction of a Hydrogel Pectin-Based Triglyceride Optical Biosensor with Immobilized Lipase Enzymes

A novel and simple optical biosensor to detect triglycerides (TGs) has been successfully constructed by using pectin hydrogel membrane as the indicator pH and chromoionophore ETH 5294 (CI), with lipase as the catalyst. The enzymatic working system against TGs releasing H+ ions will affect the color absorbance of CI. The characterization results show that a TG biosensor has the optimum condition and sensitivity at the phosphate buffer concentration of 50 mM, pH 7, and enzyme loading of 60 μg. The biosensor works at the tripalmitin (TP) concentration range of 100–400 mg/dL. With the sensitivity of 0.001 (∆A/(mg/dL)), the biosensor response reaches stability after five minutes, and the limit of detection (LOD) of the TG optical biosensor is 15 mg/dL. Relative standard deviation (RSD) in a reproducibility test was 2.5%, with a 15-day lifespan.

Wavelet Solution for Biosensor Response at Mixed Enzyme Kinetics

This paper presents the approximate solution of the reaction diffusion equation based on the hybridization of classical polynomials and Legendre wavelets. The systems of equations are generated first for the differential equations by the properties of Legendre wavelets. Theoretical analysis for the proposed scheme is discussed and computed solutions are also compared with other numerical solutions available in the literature.

Inhibitive Determination of Hg(II) in Aqueous Solution Using Urease Amperometric Biosensor

An amperometric biosensor for the indirect determination of Hg(II) has been developed based on inhibition of urease (EC 3.5.1.5) immobilized into alginate–chitosan polyelectrolyte complexes membrane. The biosensor response was monitored by following the reduction peak of hydrolyzed urea at around -0.15 V. The amperometric biosensor has a dynamic range 40–90 ppb Hg(II) with limit of detection of 66.45 ppb toward Hg(II) ions, repeatability (CV) value of 0.86% and only Ag(I) as the main potential interference. The sensor shows a stable and reproducible response for more than 2 weeks when it stored dry at 4 °C. The analytical results of Hg(II)-spiked water sample showed a good agreement with those obtained by atomic absorption spectrometry method, suggesting that the developed method may be applied in the determination of Hg(II) in the water samples.