Surface Plasmon Resonance Immunosensor for Direct Detection of Antibodies against SARS-CoV-2 Nucleocapsid Protein.

The strong immunogenicity of the SARS-CoV-2 nucleocapsid protein is widely recognized, and the detection of specific antibodies is critical for COVID-19 diagnostics in patients. This research proposed direct, label-free, and sensitive detection of antibodies against the SARS-CoV-2 nucleocapsid protein (anti-SCoV2-rN). Recombinant SARS-CoV-2 nucleocapsid protein (SCoV2-rN) was immobilized by carbodiimide chemistry on an SPR sensor chip coated with a self-assembled monolayer of 11-mercaptoundecanoic acid. When immobilized under optimal conditions, a SCoV2-rN surface mass concentration of 3.61 ± 0.52 ng/mm2 was achieved, maximizing the effectiveness of the immunosensor for the anti-SCoV2-rN determination. The calculated KD value of 6.49 × 10-8 ± 5.3 × 10-9 M confirmed the good affinity of the used monoclonal anti-SCoV2-rN antibodies. The linear range of the developed immunosensor was from 0.5 to 50 nM of anti-SCoV2-rN, where the limit of detection and the limit of quantification values were 0.057 and 0.19 nM, respectively. The immunosensor exhibited good reproducibility and specificity. In addition, the developed immunosensor is suitable for multiple anti-SCoV2-rN antibody detections.

The effect of gold nanostructure morphology on label-free electrochemical immunosensor design.

In this research, various electrodeposition techniques were used to form gold nanostructures (AuNSs) on the surface of graphite rod electrode (GE). Three distinct AuNS morphologies on GE have been achieved based on the composition of electrodeposition solution. The use of H2SO4 as a supporting electrolyte resulted in the formation of smaller but more numerous AuNSI with a modified electrode's electroactive surface area (EASA) of 0.213 cm2. Exchanging the supporting electrolyte to KNO3 and increasing HAuCl4 concentration facilitated the formation of bigger AuNSII particles with electrode EASA of 0.116 cm2. Finally, a partial coverage of GE by branched gold nanostructures (AuNSIII) was achieved with an estimated EASA of 0.110 cm2, when the HAuCl4 and KNO3 concentrations were increased further. Estimated values of heterogeneous electron transfer rate constant did not depend on AuNS morphology. Electrode modified with AuNSI exhibited the highest bovine serum albumin (BSA) immobilization efficiency and the highest relative response for the detection of specific polyclonal antibodies against BSA (p-anti-BSA) compared to other modified electrodes. The limit of p-anti-BSA detection in PBS buffer was calculated as 0.63 nM, while in blood serum it was 0.71 nM. Linear ranges were from 1 to 7 nM and from 1 to 5 nM, respectively.

ZnO nanostructures: A promising frontier in immunosensor development.

This review addresses the design of immunosensors, which employ ZnO nanostructures. Various methods of modifying ZnO nanostructures with antibodies or antigens are discussed, including covalent and non-covalent approaches and cross-linking techniques. Immunosensors based on different properties of ZnO nanomaterials are described and compared. This article provides a comprehensive review of electrochemical immunosensors based on ZnO nanostructures and various detection techniques, including cyclic voltammetry (CV), differential pulse voltammetry (DPV), photoelectrochemical (PEC) detection, electrochemical impedance spectroscopy (EIS), and other electrochemical methods. In addition, this review article examines the application of optical detection techniques, including photoluminescence (PL) and electrochemiluminescence (ECL), in the development of immunosensors based on ZnO nanostructures.

The Development of Reagentless Amperometric Glucose Biosensor Based on Gold Nanostructures, Prussian Blue and Glucose Oxidase.

Precise blood glucose detection plays a crucial role in diagnosing and medicating diabetes, in addition to aiding diabetic patients in effectively managing their condition. In this research, a first-generation reagentless amperometric glucose biosensor was developed by combining the graphite rod (GR) electrode modification by gold nanostructures (AuNS) and Prussian blue (PB) with glucose oxidase (GOx)-an enzyme that can oxidize glucose and produce H2O2. Firstly, AuNS was electrochemically deposited on the GR electrode (AuNS/GR), and then PB was electrochemically synthesized on the AuNS/GR electrode (PB/AuNS/GR). Finally, GOx was immobilized over the PB/AuNS nanocomposite with the assistance of Nafion (Nf) (Nf-GOx/PB/AuNS/GR). An application of PB in the design of a glucose biosensor enables an easy electrochemical reduction and, thus, the determination of the H2O2 produced during the GOx-catalyzed oxidation of glucose in the sample at a low operation potential of -0.05 V vs. Ag/AgCl/KCl3 mol L-1. In addition, AuNS increased the electrochemically active surface area, improved the GOx immobilization and ensured a higher analytical signal. The developed glucose biosensor based on the Nf-GOx/PB/AuNS/GR electrode exhibited a wide linear range, from 0.025 to 1 mmol L-1 of glucose, with a 0.0088 mmol L-1 limit of detection, good repeatability and high selectivity over electroactive interfering substances. The developed biosensor is convenient for the determination of glucose in the physiological environment.

Electrochemical Immunosensor for the Determination of Antibodies against Prostate-Specific Antigen Based on ZnO Nanostructures.

In this study, ZnO nanostructures with different types of morphologies and particle sizes were evaluated and applied for the development of an immunosensor. The first material was composed of spherical, polydisperse nanostructures with a particle size in the range of 10-160 nm. The second was made up of more compact rod-like spherical nanostructures with the diameter of these rods in the range of 50-400 nm, and approximately 98% of the particles were in the range of 20-70 nm. The last sample of ZnO was made up of rod-shaped particles with a diameter of 10-80 nm. These ZnO nanostructures were mixed with Nafion solution and drop-casted onto screen-printed carbon electrodes (SPCE), followed by a further immobilization of the prostate-specific antigen (PSA). The affinity interaction of PSA with monoclonal antibodies against PSA (anti-PSA) was evaluated using the differential pulse voltammetry technique. The limit of detection and limit of quantification of anti-PSA were determined as 1.35 nM and 4.08 nM for compact rod-shaped spherical ZnO nanostructures, and 2.36 nM and 7.15 nM for rod-shaped ZnO nanostructures, respectively.

Magneto-Immunoassay for the Detection and Quantification of Human Growth Hormone.

Physiological and endocrine maintenance of a normal human growth hormone (hGH) concentration is crucial for growth, development, and a number of essential biological processes. In this study, we describe the preparation and characterization of magnetic nanoparticles coated with a gold shell (MNPs-Au). The optimal surface concentration of monoclonal anti-hGH antibodies (m-anti-hGH) on magnetic nanoparticles, as well as conditions that decrease non-specific interactions during the magneto-immunoassay, were elaborated. After the selective recognition, separation, and pre-concentration of hGH by MNPs-Au/m-anti-hGH and the hGH interaction with specific polyclonal biotin-labeled antibodies (p-anti-hHG-B) and streptavidin modified horseradish peroxidase (S-HRP), the MNPs-Au/m-anti-hGH/hGH/p-anti-hGH-B/S-HRP immunoconjugate was formed. The concentration of hGH was determined after the addition of 3,3',5,5'-tetramethylbenzidine and hydrogen peroxide substrate solution for HRP; the absorbance at 450 nm was registered after the addition of STOP solution. The developed sandwich-type colorimetric magneto-immunoassay is characterized by a clinically relevant linear range (from 0.1 to 5.0 nmol L-1, R2 0.9831), low limit of detection (0.082 nmol L-1), and negligible non-specific binding of other antibodies or S-HRP. The obtained results demonstrate the applicability of the developed magneto-immunoassay for the concentration and determination of hGH in the serum. Additionally, important technical solutions for the development of the sandwich-type colorimetric magneto-immunoassay are discussed.

Gold nanoparticle based colorimetric sensing strategy for the determination of reducing sugars.

The colorimetric sensors for reducing sugars based on a redox reaction between AuCl4- ions and fructose, glucose, lactose, or mannose are presented. Gold nanoparticles (AuNPs) that formed at room temperature as a product of this reaction were registered using a spectrophotometer. Lengthening reaction time had a positive effect on the sensitivity of the developed sensors. Different reducing sugars exhibited distinct reaction rates for AuNP formation, with the rate decreasing in the order fructose > glucose > lactose > mannose. LOD values after 60 min of the reaction for different sugars followed the same trend of 0.067, 0.081, 0.087, and 0.106 mM, while LOQ was 0.223, 0.270, 0.289, and 0.353 mM, respectively. The linear range 60 min since the start of the reaction varied from 0.3 up to 5.0 mM for different sugars. The colorimetric sensor was evaluated for use in real samples of beverages, milk, and saliva.

Electrodeposited Gold Nanostructures for the Enhancement of Electrochromic Properties of PANI-PEDOT Film Deposited on Transparent Electrode.

Conjugated polymers (CPs) are attractive materials for use in different areas; nevertheless, the enhancement of electrochromic stability and switching time is still necessary to expand the commercialization of electrochromic devices. To our best knowledge, this is the first study demonstrating the employment of electrodeposited gold nanostructures (AuNS) for the enhancement of CPs' electrochromic properties when a transparent electrode is used as a substrate. Polyaniline-poly(3,4-ethylenedioxythiophene) (PANI-PEDOT) films were electrodeposited on a transparent indium tin oxide glass electrode, which was pre-modified by two different methods. AuNS were electrodeposited at -0.2 V constant potential for 60 s using both the 1st method (synthesis solution consisted of 3 mM HAuCl4 and 0.1 M H2SO4) and 2nd method (15 mM HAuCl4 and 1 M KNO3) resulting in an improvement of optical contrast by 3% and 22%, respectively. Additionally, when using the 1st method, the coloration efficiency was improved by 50% while the switching time was reduced by 17%. Furthermore, in both cases, the employment of AuNS resulted in an enhancement of the electrochromic stability of the CPs layer. A further selection of AuNS pre-modification conditions with the aim to control their morphology and size can be a possible stepping stone for the further improvement of CPs electrochromic properties.