This article aims to present a comparative study of three polypyrrole-based molecularly imprinted polymer (MIP) systems for the detection of the recombinant severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (rN). The rN is known for its relatively low propensity to mutate compared to other SARS-CoV-2 antigens. The aforementioned systems include screen-printed carbon electrodes (SPCE) modified with gold nanostructures (MIP1), platinum nanostructures (MIP2), and the unmodified SPCE (MIP3), which was used for control. Pulsed amperometric detection (PAD) was employed as the detection technique, offering the advantage of label-free detection without the need for an additional redox probe. Calibration curves were constructed using the obtained data to evaluate the response of each system. Non-imprinted systems were also tested in parallel to evaluate the contribution of non-specific binding and assess the affinity sensor\'s efficiency. The analysis of calibration curves revealed that the AuNS-based MIP1 system exhibited the lowest contribution of non-specific binding and displayed a better fit with the chosen fitting model compared to the other systems. Further analysis of this system included determining the limit of detection (LOD) (51.2 ± 2.8 pg/mL), the limit of quantification (LOQ) (153.9 ± 8.3 pg/mL), and a specificity test using a recombinant receptor-binding domain of SARS-CoV-2 spike protein as a control. Based on the results, the AuNS-based MIP1 system demonstrated high specificity and sensitivity for the label-free detection of SARS-CoV-2 nucleocapsid protein. The utilization of PAD without the need for additional redox probes makes this sensing system convenient and valuable for rapid and accurate virus detection.

Affinity biosensors play a crucial role in clinical diagnosis, pharmaceuticals, immunology, and other areas of human health. Affinity biosensors rely on the specific binding between target analytes and biological ligands such as antibodies, nucleic acids, aptamers, or other receptors to primarily generate electrochemical or optical signals. Considerable effort has been put into improving the performance of the affinity technologies to make them more sensitive, efficient and reproducible, of the many approaches electrokinetic phenomena are a viable option. In this perspective, studies that combine electrokinetic phenomena with affinity biosensor are discussed about their promise for achieving higher sensitivity and lower detection limit.

Roadside testing of illicit drugs such as tetrahydrocannabinol (THC) requires simple, rapid, and cost-effective methods. The need for non-invasive detection tools has led to the development of selective and sensitive platforms, able to detect phyto- and synthetic cannabinoids by means of their main metabolites in breath, saliva, and urine samples. One may estimate the time passed from drug exposure and the frequency of use by corroborating the detection results with pharmacokinetic data. In this review, we report on the current detection methods of cannabinoids in biofluids. Fluorescent, electrochemical, colorimetric, and magnetoresistive biosensors will be briefly overviewed, putting emphasis on the affinity formats amenable to on-site screening, with possible applications in roadside testing and anti-doping control.

Biosensors play a central role in moving diagnostics to being on-site or decentralized. Affinity biosensor, an important category of biosensors, has important applications in clinical diagnosis, pharmaceuticals, immunology, and other fields. Affinity biosensors rely on specific binding between target analytes and biological ligands such as antibodies, nucleic acids, or other receptors to generate measurable signals. Oftentimes the target analytes in practical samples are of low abundance in a complex matrix. Traditional affinity biosensors mainly rely on random diffusion of analytes in solution to conjugate with biorecognition elements on the sensor surface of electrodes. The process may take hours or even days, which is not conducive to rapid and sensitive detection of biosensors. Therefore, it is strongly desired to incorporate an enrichment mechanism for target analytes into biosensor-based detection. AC electrokinetic (ACEK) effect can realize rapid enrichment of analytes by application of AC electric fields, which holds great promise for achieving high sensitivity, low detection limit, and rapid turnaround. This article reviews the studies of affinity biosensors integrated with ACEK enrichment in the past decade, and summarizes the latest detection methods, detection devices and applications, hoping to provide some insights and references for researchers in related fields.

Point-of-care (POC) testing has revolutionized diagnostic healthcare, bringing medical results directly and immediately to the patient. With faster diagnostics, more immediate clinical management decisions can be made. POC tests most often use a dipstick or swab format to detect the presence of a pathogen, disease, or other relevant biomarker. In these formats, the POC tests eliminate the need for complex lab equipment and trained personnel to collect, process, and analyze sample data for simple diagnostics. However, these tests cannot satisfy all clinical needs, because accurate quantitative results are needed. The present study serves as a template for designing a nonfaradaic electrochemical biosensor toward quantitative POC diagnostics. We focus on investigating the most important parameters when constructing a nonfaradaic biosensor through both mathematical modeling and electrochemical measurements. Furthermore, we demonstrate quantitative affinity biosensing of a model protein toward developing a POC device.

In spite of the high analytic potential of Magneto Optical Surface Plasmon Resonance (MOSPR) assays, their applicability to biosensing has been limited due to significant chip stability issues. We present novel solutions to surpass current limitations of MOSPR sensing assays, based on innovative chip structure, tailored measurements and improved data analysis methods. The structure of the chip is modified to contain a thin layer of Co-Au alloy instead of successive layers of homogenous metals with magnetic and plasmonic properties, as currently used. This new approach presents improved plasmonic and magnetic properties, yet a structural stability similar to standard Au-SPR chips, allowing for bioaffinity assays in saline solutions. Moreover, using a custom-designed measurement configuration that allows the acquisition of the SPR curve, i.e., the reflectivity measured at multiple angles of incidence, instead of the reflectivity value at a single-incidence angle, a high signal-to-noise ratio is achieved, suitable for detection of minute analyte concentrations. The proposed structure of the MOSPR sensing chip and the procedure of data analysis allow for long time assessment in liquid media, a significant advancement over existing MOSPR chips, and confirm the MOSPR increased sensitivity over standard SPR analyses.

The developed surface plasmon resonance (SPR) biosensor based on the recombinant Staphylococcal protein A with an additional cysteine residue (SPA-Cys) used as a biorecognition component showed a good selectivity and sensitivity for the immunoglobulin detection. The developed biosensor with SPA-Cys-based bioselective element can also be used as a first step of immunosensor creation. The successful immobilization of SPA-Cys on the nanolayer gold sensor surface of the SPR spectrometer was performed. The efficiency of blocking nonspecific sorption sites on the sensor surface with milk proteins, gelatin, BSA, and HSA was studied, and a rather high efficiency of using gelatin was confirmed. The SPR biosensor selectively interacted with IgG and did not interact with the control proteins. The linear dependence of the sensor response on the IgG concentration in the range from 2 to 10 μg/ml was shown. Using the calibration curve, the IgG concentration was measured in the model samples. The determined concentrations are in good agreement (r 2 = 0.97) with the given concentration of IgG.

Biosensors are a very active research field. They have the potential to lead to low-cost, rapid, sensitive, reproducible, and miniaturized bioanalytical devices, which exploit the high binding avidity and selectivity of biospecific binding molecules together with highly sensitive detection principles. Of the optical biosensors, those based on chemical luminescence detection (including chemiluminescence, bioluminescence, electrogenerated chemiluminescence, and thermochemiluminescence) are particularly attractive, due to their high-to-signal ratio and the simplicity of the required measurement equipment. Several biosensors based on chemical luminescence have been described for quantitative, and in some cases multiplex, analysis of organic molecules (such as hormones, drugs, pollutants), proteins, and nucleic acids. These exploit a variety of miniaturized analytical formats, such as microfluidics, microarrays, paper-based analytical devices, and whole-cell biosensors. Nevertheless, despite the high analytical performances described in the literature, the field of chemical luminescence biosensors has yet to demonstrate commercial success. This review presents the main recent advances in the field and discusses the approaches, challenges, and open issues, with the aim of stimulating a broader interest in developing chemical luminescence biosensors and improving their commercial exploitation.

We present an electrochemical and optical characterization of 5,10,15,20-tetraphenylporphyrin tin(IV) dichloride (Sn-tpp) in terms of its potential use as a hybrid proteins\' label. Our research comprised Sn-tpp and Sn-tpp in the presence of model proteins selected as to mimic a receptor or surface blocking agents: bovine serum albumin, ovalbumin, and immunoglobulin G. In the course of the study, we determined optimal conditions for analysis by means of differential pulse voltammetry, ultraviolet-visible spectrophotometry, and spectrofluorimetry. In electrochemical detection, the influence of the working electrode, solvent, and supporting electrolyte was examined. Displacements of the received signals along the potential axis (a shift of the potential) and changes in signal intensities due to the addition of proteins were observed and analyzed. Simultaneously, the suitability of Sn-tpp as a label in optical detection mode was assessed by using spectroscopic techniques. The obtained results prove Sn-tpp to be applicable in dual and triple detection systems. Such an approach will improve the reliability of the analysis and, at the same time, will allow for widening the range of the linear response with some overlapping ranges of concentrations.

We present novel solutions to surpass current analytic limitations of Magneto-Optical Surface Plasmon Resonance (MOSPR) assays, concerning both the chip structure and the method for data analysis. The structure of the chip is modified to contain a thin layer of Co-Au alloy instead of successive layers of homogeneous metals, as currently used. This alloy presents improved plasmonic and magnetic properties, yet a structural stability similar to Au-SPR chips, allowing for bioaffinity assays in saline solutions. Analyzing the whole reflectivity curve at multiple angles of incidence instead of the reflectivity value at a single incidence angle provides a high signal-to-noise ratio suitable for detection of minute analyte concentrations. Based on assessment of solutions with known refractive indices as well as of a model biomolecular interaction (i.e. IgG-AntiIgG) we demonstrate that the proposed structure of the MOSPR sensing chip and the procedure of data analysis allows for long-time assessment in liquid media with increased sensitivity over standard SPR analyses.