MIP-based electrochemical sensor for direct detection of hepatitis C virus via E2 envelope protein.

Hepatitis C is the most common liver disease caused by Hepatitis C virus (HCV), and can evolve into serious health problems e.g. cirrhosis and hepatocellular carcinoma. Nowadays, the initial stage of the disease cannot be practically diagnosed, representing thus an extremely important problem of modern public health care. This study is aimed at the development of a sensor for direct detection of HCV. The sensor utilizes a synthetic recognition element prepared by the technology of molecular imprinting and representing a molecularly imprinted polymer (MIP) having molecular recognition sites of HCV envelope protein E2 (E2-MIP). E2-MIP integrated into an electrochemical sensor platform allows quantitative evaluation of binding of free E2 protein as well as HCV-mimetic particles (HCV-MPs) in human plasma with LOD value of 4.6 × 10-4 ng/mL (for HCV-MPs). The developed electrochemical HCV sensor represents a simple, fast and inexpensive alternative for the existing methods of HCV detection and paves the way for the point-of care diagnostics of Hepatitis C.

Molecularly imprinted polymer based electrochemical sensor for quantitative detection of SARS-CoV-2 spike protein.

The continued spread of the coronavirus disease and prevalence of the global pandemic is exacerbated by the increase in the number of asymptomatic individuals who unknowingly spread the SARS-CoV-2 virus. Although remarkable progress is being achieved at curtailing further rampage of the disease, there is still the demand for simple and rapid diagnostic tools for early detection of the COVID-19 infection and the following isolation. We report the fabrication of an electrochemical sensor based on a molecularly imprinted polymer synthetic receptor for the quantitative detection of SARS-CoV-2 spike protein subunit S1 (ncovS1), by harnessing the covalent interaction between 1,2-diols of the highly glycosylated protein and the boronic acid group of 3-aminophenylboronic acid (APBA). The sensor displays a satisfactory performance with a reaction time of 15 min and is capable of detecting ncovS1 both in phosphate buffered saline and patient's nasopharyngeal samples with LOD values of 15 fM and 64 fM, respectively. Moreover, the sensor is compatible with portable potentiostats thus allowing on-site measurements thereby holding a great potential as a point-of-care testing platform for rapid and early diagnosis of COVID-19 patients.

An electrochemical biosensor for direct detection of hepatitis C virus.

This paper is aimed at the development of a biosensor for direct detection of Hepatitis C virus (HCV) surface antigen: envelope protein (E2). A recombinant LEL fragment of biological cell receptor CD81 and two short synthetic peptides imitating the fragment of LEL sequence of CD81 (linear and loop-like peptides) capable of specific binding to E2 were tested as molecular recognition elements of the biosensor. For this purpose the selected ligands were immobilized to the surface of a screen-printed electrode utilized as an electrochemical sensor platform. The immobilization parameters such as the ligand concentration and the immobilization time were carefully optimized for each ligand. Differential pulse voltammetry used to evaluate quantitatively binding of E2 to the ligands revealed their similar binding affinity towards E2. Thus, the linear peptide was selected as a less expensive and easily prepared ligand for the HCV biosensor preparation. The resulting HCV biosensor demonstrated selectivity towards E2 in the presence of interfering protein, conalbumin. Moreover, it was found that the prepared biosensor effectively detected E2 bound to hepatitis C virus-mimetic particles (HC VMPs) at LOD value of 2.1∙10-5 mg/mL both in 0.01 M PBS solution (pH 7.4) and in simulated blood plasma.

Development of a portable MIP-based electrochemical sensor for detection of SARS-CoV-2 antigen.

The current COVID-19 pandemic caused by SARS-CoV-2 coronavirus is expanding around the globe. Hence, accurate and cheap portable sensors are crucially important for the clinical diagnosis of COVID-19. Molecularly imprinted polymers (MIPs) as robust synthetic molecular recognition materials with antibody-like ability to bind and discriminate between molecules can perfectly serve in building selective elements in such sensors. Herein, we report for the first time on the development of a MIP-based electrochemical sensor for detection of SARS-CoV-2 nucleoprotein (ncovNP). A key element of the sensor is a disposable sensor chip - thin film electrode - interfaced with a MIP-endowed selectivity for ncovNP and connected with a portable potentiostat. The resulting ncovNP sensor showed a linear response to ncovNP in the lysis buffer up to 111 fM with a detection and quantification limit of 15 fM and 50 fM, respectively. Notably, the sensor was capable of signaling ncovNP presence in nasopharyngeal swab samples of COVID-19 positive patients. The presented strategy unlocks a new route for the development of rapid COVID-19 diagnostic tools.

Molecularly imprinted polymer-based sensor for electrochemical detection of erythromycin.

The increasing global reports on the occurrence of macrolide antibiotics resistance, especially erythromycin (Ery) resistant strains, suggests the possible presence of these antibiotics in the environment hence, their inclusion in the EU watchlist of water pollutants. Consequently, there is an urgent need for the development of portable and cost effective analytical sensing devices for their monitoring in water. The combination of molecularly imprinted polymer (MIP) as a sensing element with a portable electrochemical transducer such as screen printed electrode (SPE) may offer a valuable approach for the desired routine environmental monitoring. This work demonstrates the preparation of an electrochemical MIP-based sensor for Ery detection in aqueous media. Ery-selective MIP, Ery-MIP was generated directly on SPE, Ery-MIP/SPE via electrochemical polymerization of m-phenylenediamine (mPD). The optimization of sensor performance was achieved with special attention given to the selection of functional monomer, template removal, polymer thickness and incubation time. Ery-MIP/SPE sensor demonstrated the ability to discriminate target analyte against very close analogues i.e clarithromycin and azithromycin in both PBS and tap water. In addition, Ery-MIP/SPE could detect Ery down to low limits (LOD = 0.1 nM and LOQ = 0.4 nM) and exhibited good recovery in tap water. The presented analytical approach could be potentially suited and/or further developed for adequate monitoring of Ery as well as other macrolides in environmental water.

Hybrid molecularly imprinted polymer for amoxicillin detection.

The potential adverse effects of the environmental presence of antibiotics on the ecosystem demands the development of new methods suitable for accurate detection of these micropollutants in various aquatic media. An analytical method exploiting the synergistic effect of a label-free sensing platform combined with a molecularly imprinted polymer (MIP) as robust recognition element could represent an efficient tool for the real-time monitoring of antibiotics. In this work, a hybrid organic-inorganic MIP film (AMO-MIP) selective towards amoxicillin (AMO) was synthesized and integrated with a surface plasmon resonance (SPR) sensor. The film was prepared by sol-gel using methacrylamide (MAAM) as organic functional monomer, tetraethoxysilane (TEOS) as inorganic precursor, and vinyltrimethoxysilane (VTMOS) as coupling agent. The AMO-MIP film characterized with the SPR system demonstrated about 16 times higher binding capacity to AMO than corresponding reference non-imprinted polymer (NIP). AMO-MIP-modified SPR sensors could detect AMO with LoD down to 73 pM and discriminate AMO among structurally similar molecules both in buffer and in tap water. Good reproducibility was achieved for several rebinding-regeneration cycles. The sensor could be stored at room temperature for up to 6 months without losing stability.

Molecularly Imprinted Polymer Integrated with a Surface Acoustic Wave Technique for Detection of Sulfamethizole.

The synergistic effect of combining molecular imprinting and surface acoustic wave (SAW) technologies for the selective and label-free detection of sulfamethizole as a model antibiotic in aqueous environment was demonstrated. A molecularly imprinted polymer (MIP) for sulfamethizole (SMZ) selective recognition was prepared in the form of a homogeneous thin film on the sensing surfaces of SAW chip by oxidative electropolymerization of m-phenylenediamine (mPD) in the presence of SMZ, acting as a template. Special attention was paid to the rational selection of the functional monomer using computational and spectroscopic approaches. SMZ template incorporation and its subsequent release from the polymer was supported by IR microscopic measurements. Precise control of the thicknesses of the SMZ-MIP and respective nonimprinted reference films (NIP) was achieved by correlating the electrical charge dosage during electrodeposition with spectroscopic ellipsometry measurements in order to ensure accurate interpretation of label-free responses originating from the MIP modified sensor. The fabricated SMZ-MIP films were characterized in terms of their binding affinity and selectivity toward the target by analyzing the binding kinetics recorded using the SAW system. The SMZ-MIPs had SMZ binding capacity approximately more than eight times higher than the respective NIP and were able to discriminate among structurally similar molecules, i.e., sulfanilamide and sulfadimethoxine. The presented approach for the facile integration of a sulfonamide antibiotic-sensing layer with SAW technology allowed observing the real-time binding events of the target molecule at nanomolar concentration levels and could be potentially suitable for cost-effective fabrication of a multianalyte chemosensor for analysis of hazardous pollutants in an aqueous environment.

Molecularly imprinted polymer film interfaced with Surface Acoustic Wave technology as a sensing platform for label-free protein detection.

Molecularly imprinted polymer (MIP)-based synthetic receptors integrated with Surface Acoustic Wave (SAW) sensing platform were applied for the first time for label-free protein detection. The ultrathin polymeric films with surface imprints of immunoglobulin G (IgG-MIP) were fabricated onto the multiplexed SAW chips using an electrosynthesis approach. The films were characterized by analyzing the binding kinetics recorded by SAW system. It was revealed that the capability of IgG-MIP to specifically recognize the target protein was greatly influenced by the polymer film thickness that could be easily optimized by the amount of the electrical charge consumed during the electrodeposition. The thickness-optimized IgG-MIPs demonstrated imprinting factors towards IgG in the range of 2.8-4, while their recognition efficiencies were about 4 and 10 times lower toward the interfering proteins, IgA and HSA, respectively. Additionally, IgG-MIP preserved its capability to recognize selectively the template after up to four regeneration cycles. The presented approach of the facile integration of the protein-MIP sensing layer with SAW technology allowed observing the real-time binding events of the target protein at relevant sensitivity levels and can be potentially suitable for cost effective fabrication of a biosensor for analysis of biological samples in multiplexed manner.