Imprinted Polymer Nanoparticles Research Articles

Bidirectional Regulation of Intracellular Enzyme Activity Using Light-Driven Nano-Inhibitors.

Photochemical regulation provides precise control over enzyme activities with high spatiotemporal resolution. A promising approach involves anchoring "photoswitches" at enzyme active sites to modulate substrate recognition. However, current methods often require genetic mutations and irreversible enzyme modifications for the site-specific anchoring of "photoswitches", potentially compromising the enzyme activities. Herein, we present a pioneering reversible nano-inhibitor based on molecular imprinting technique for bidirectional regulation of intracellular enzyme activity. The nano-inhibitor employs a molecularly imprinted polymer nanoparticle as its body and azobenzene-modified inhibitors ("photoswitches") as the arms. By using a target enzyme as the molecular template, the nano-inhibitor acquires oriented binding sites on its surface, resulting in a high affinity for the target enzyme and non-covalently firm anchoring of the azobenzene-modified inhibitor to the enzyme active site. Harnessing the reversible isomerization of azobenzene units upon exposure to ultraviolet and visible light, the nano-inhibitor achieves bidirectional enzyme activity regulation by precisely docking and undocking inhibitor at the active site. Notably, this innovative approach enables the facile in situ regulation of intracellular endogenous enzymes, such as carbonic anhydrase. Our results represent a practical and versatile tool for precise enzyme activity regulation in complex intracellular environments.

Bidirectional Regulation of Intracellular Enzyme Activity Using Light‐Driven Nano‐Inhibitors

AbstractPhotochemical regulation provides precise control over enzyme activities with high spatiotemporal resolution. A promising approach involves anchoring “photoswitches” at enzyme active sites to modulate substrate recognition. However, current methods often require genetic mutations and irreversible enzyme modifications for the site‐specific anchoring of “photoswitches”, potentially compromising the enzyme activities. Herein, we present a pioneering reversible nano‐inhibitor based on molecular imprinting technique for bidirectional regulation of intracellular enzyme activity. The nano‐inhibitor employs a molecularly imprinted polymer nanoparticle as its body and azobenzene‐modified inhibitors (“photoswitches”) as the arms. By using a target enzyme as the molecular template, the nano‐inhibitor acquires oriented binding sites on its surface, resulting in a high affinity for the target enzyme and non‐covalently firm anchoring of the azobenzene‐modified inhibitor to the enzyme active site. Harnessing the reversible isomerization of azobenzene units upon exposure to ultraviolet and visible light, the nano‐inhibitor achieves bidirectional enzyme activity regulation by precisely docking and undocking inhibitor at the active site. Notably, this innovative approach enables the facile in situ regulation of intracellular endogenous enzymes, such as carbonic anhydrase. Our results represent a practical and versatile tool for precise enzyme activity regulation in complex intracellular environments.

Molecularly imprinted polymer nanoparticle-carbon nanotube composite electrochemical gas sensor for highly selective and sensitive detection of methanol vapour.

An electrochemical gas sensor has been fabricated using molecularly imprinted polymer nanoparticles (nanoMIPs) and multiwalled carbon nanotubes on screen-printed electrodes. Methanol vapour was chosen as the target due to its toxicity as its suitability as a model for more harmful pollutant gases. The sensor functions under ambient conditions and in the required concentration range, in contrast to all previous MIP-based gas sensors for methanol. The sensitivity of the sensor was greatly improved by the addition of multiwall carbon nanotubes, resulting in a limit of detection of approximately 10 ppm. The nanoMIPs provide an inherent selectivity for the target inherent in its design. Selectivity studies were performed with structurally analogous alcohols at various concentrations, demonstrating selectivity for methanol 12.1 times that for ethanol at 2 mmol dm-3 and 4.2 times that for ethanol at 1 mmol dm-3. Interactions with isopropanol and n-propanol were found to be non-specific, and the response to water was negligible. This demonstrates an improvement over previous methanol gas sensors based on molecularly imprinted polymers. No response was observed with carbon nanotubes alone, and no selectivity was observed with non-imprinted equivalents of the nanoMIP sensor. The resulting device is by far the most practical MIP-based instrument for methanol gas sensing thus far described in the literature, being the only example capable of functioning at the necessary methanol vapour concentrations and at the required temperature and humidity. With the selectivity and sensitivity described and the simple design, the developed device provides a substantial advance in the field of molecularly imprinted gas sensors.

Preparation of molecularly imprinted polymer nanoparticles for selective adsorption of caffeine using dual-functionalized Ag2S quantum dots

Caffeine is the most abundant and widely consumed active medicinal compound, making it the most representative pollutant in the environment. This research involved synthesizing high-performance fluorescent HOOC-CC-functionalized Ag2S quantum dots (HOOC-QDs) measuring approximately 4.35 nm in size using the hydrothermal method. These QDs were then used to produce fluorescent molecularly imprinted polymer nanoparticles (MIP-Ag2S NPs) measuring 20.31 nm. The MIP-Ag2S NPs were created through a graft suspension copolymerization process involving methacrylic acid monomer, ethylene glycol dimethacrylate cross-linker, and caffeine template molecules. The polar carboxylic acid groups enhance the interaction with the template molecules, which is crucial for selectivity, and the -CC- groups aid in network formation. The MIP-Ag2S NPs emitted fluorescence at around 500 nm, which was significantly reduced after caffeine adsorption. The pH-sensitive MIP-Ag2S NPs had a maximum caffeine adsorption capacity of 488 mg/g at pH 6 and a maximum release of approximately 96% at pH 2.2. Additionally, repeated cycles of adsorption and desorption did not result in a significant loss of adsorption capacity. The HOOC-QDs can be used to prepare MIPs for extracting and recognizing pollutants and medicine in water and for controlled drug delivery systems.

Modulation of EGFR Activity by Molecularly Imprinted Polymer Nanoparticles Targeting Intracellular Epitopes.

In recent years, molecularly imprinted polymer nanoparticles (nanoMIPs) have proven to be an attractive alternative to antibodies in diagnostic and therapeutic applications. However, several key questions remain: how suitable are intracellular epitopes as targets for nanoMIP binding? And to what extent can protein function be modulated via targeting specific epitopes? To investigate this, three extracellular and three intracellular epitopes of epidermal growth factor receptor (EGFR) were used as templates for the synthesis of nanoMIPs which were then used to treat cancer cells with different expression levels of EGFR. It was observed that nanoMIPs imprinted with epitopes from the intracellular kinase domain and the extracellular ligand binding domain of EGFR caused cells to form large foci of EGFR sequestered away from the cell surface, caused a reduction in autophosphorylation, and demonstrated effects on cell viability. Collectively, this suggests that intracellular domain-targeting nanoMIPs can be a potential new tool for cancer therapy.

Molecularly Imprinted Nanomedicine for Anti-angiogenic Cancer Therapy via Blocking Vascular Endothelial Growth Factor Signaling.

The VEGF-VEGFR2 (VEGF = vascular endothelial growth factor) signaling has been a promising target in cancer therapy. However, because conventional anti-angiogenic therapeutics suffer from drawbacks, particularly severe side effects, novel anti-angiogenic strategies are much needed. Herein, we report the rational engineering of VEGF-targeted molecularly imprinted polymer nanoparticles (nanoMIP) for anti-angiogenic cancer therapy. The anti-VEGF nanomedicine was prepared via a state-of-the-art molecular imprinting approach using the N-terminal epitope of VEGF as the template. The nanoMIP could target the two major pro-angiogenic isoforms (VEGF165 and VEGF121) with high affinity and thereby effectively block the VEGF-VEGFR2 signaling, yielding a potent anti-angiogenic effect of "killing two birds with one stone". In vivo experiments demonstrated that the anti-VEGF nanoMIP effectively suppressed tumor growth via anti-angiogenesis in a xenograft model of human colon carcinoma without apparent side effects. Thus, this study not only proposes an unprecedented anti-angiogenic strategy for cancer therapy but also provides a new paradigm for the rational development of MIPs-based "drug-free" nanomedicines.

Development of Molecularly Imprinted Polymer Nanoparticles with High Affinity and Its Application to Electrochemical Biosensors

Molecularly imprinted polymers (MIPs) have the potentials to be low-cost and stable alternatives to natural receptors such as enzymes and antibodies for biosensors. In our group, we have been developing enzyme-/antibody-free electrochemical biosensors with the MIP-coated electrodes [1,2]. However, the conventional polymerization methods for MIPs have the limitations such as their heterogeneities that reduce high binding affinity sites and the presence of residual templated molecules, which are detrimental to biomolecular recognition. To address these issues, we have synthesized molecularly imprinted polymer nanoparticles (nano-MIPs) with virtually free of heterogeneous templates and higher affinity for target biomolecules by using a solid-phase approach synthesis method [3]. In this study, we have focused on the biomarkers such as human serum albumin (HSA) and glycated albumin (GA) for diabetes and then synthesized the redox-labeled HSA-templated nano-MIPs (HNMs) and GA-templated nano-MIPs (GNMs) for the electrochemical biosensor (Figure 1). The fundamental characteristics of HNMs and GNMs such as chemical composition, particle size, and binding affinity have been evaluated by using proton nuclear magnetic resonance (1H NMR), dynamic light scattering (DLS), atomic force microscopy (AFM), cyclic voltammetry (CV), isothermal titration calorimetry (ITC), and surface plasmon resonance (SPR).References Kajisa, T.; Sakata, T. ACS Appl. Mater. Interfaces 2018,10, 34983–34990.Sakata, T.; Nishitani, S.; Kajisa, T. RSC Advances 2020, 10, 16999–17013.Canfarotta, F. et al. Nat. Protoc. 2016, 3, 443-455. Figure 1

Highly selective molecularly imprinted polymer nanoparticles (MIP NPs)-based microfluidic gas sensor for tetrahydrocannabinol (THC) detection

A highly selective microfluidic integrated metal oxide gas sensor for THC detection is reported based on MIP nanoparticles (MIP NPs). We synthesized MIP NPs with THC recognition sites and coated them on a 3D-printed microfluidic channel surface. The sensitivity and selectivity of coated microfluidic integrated gas sensors were evaluated by exposure to THC, cannabidiol (CBD), methanol, and ethanol analytes in 300–700 ppm at 300 °C. For comparison, reference signals were obtained from a microfluidic channel coated with nonimprinted polymers (NIP NPs). The MIP and NIP NPs were characterized using scanning electron microscopy (SEM) and Raman spectroscopy. MIP and NIP NPs channels response data were combined and classified with 96.3% accuracy using the Fine KNN classification model in MATLAB R2021b Classification Learner App. Compared to the MIP NPs coated channel, the NIP NPs channel had poor selectivity towards THC, demonstrating that the THC recognition sites in the MIP structure enabled selective detection of THC. The findings demonstrated that the recognition sites of MIP NPs properly captured THC molecules, enabling the selective detection of THC compared to CBD, methanol, and ethanol.

Imprinted Hydrogel Nanoparticles for Protein Biosensing: A Review.

Over the past decade, molecular imprinting (MI) technology has made tremendous progress, and the advancements in nanotechnology have been the major driving force behind the improvement of MI technology. The preparation of nanoscale imprinted materials, i.e., molecularly imprinted polymer nanoparticles (MIP NPs, also commonly called nanoMIPs), opened new horizons in terms of practical applications, including in the field of sensors. Currently, hydrogels are very promising for applications in bioanalytical assays and sensors due to their high biocompatibility and possibility to tune chemical composition, size (microgels, nanogels, etc.), and format (nanostructures, MIP film, fibers, etc.) to prepare optimized analyte-responsive imprinted materials. This review aims to highlight the recent progress on the use of hydrogel MIP NPs for biosensing purposes over the past decade, mainly focusing on their incorporation on sensing devices for detection of a fundamental class of biomolecules, the peptides and proteins. The review begins by directing its focus on the ability of MIPs to replace biological antibodies in (bio)analytical assays and highlight their great potential to face the current demands of chemical sensing in several fields, such as disease diagnosis, food safety, environmental monitoring, among others. After that, we address the general advantages of nanosized MIPs over macro/micro-MIP materials, such as higher affinity toward target analytes and improved binding kinetics. Then, we provide a general overview on hydrogel properties and their great advantages for applications in the field of Sensors, followed by a brief description on current popular routes for synthesis of imprinted hydrogel nanospheres targeting large biomolecules, namely precipitation polymerization and solid-phase synthesis, along with fruitful combination with epitope imprinting as reliable approaches for developing optimized protein-imprinted materials. In the second part of the review, we have provided the state of the art on the application of MIP nanogels for screening macromolecules with sensors having different transduction modes (optical, electrochemical, thermal, etc.) and design formats for single use, reusable, continuous monitoring, and even multiple analyte detection in specialized laboratories or in situ using mobile technology. Finally, we explore aspects about the development of this technology and its applications and discuss areas of future growth.

Electroactive molecularly imprinted polymer nanoparticles for selective glyphosate determination

Redox-active molecularly imprinted polymer nanoparticles selective for glyphosate, MIP-Gly NPs, were devised, synthesized, and subsequently integrated onto platinum screen-printed electrodes (Pt-SPEs) to fabricate a chemosensor for selective determination of glyphosate (Gly) without the need for redox probe in the test solution. That was because, ferrocenylmethyl methacrylate was added to the polymerization mixtures during the NPs synthesis so that the resulting MIP-Gly NPs contained covalently immobilized ferrocenyl moieties as the reporting redox ingredient, conferring these NPs with electroactive properties. MIP-Gly NPs of four different compositions were evaluated. The herein described approach represents a simple and effective way to endow MIP NPs with electrochemical reporting capabilities with neither the need to functionalize them post-synthesis nor to use electrochemical mediators present in the tested solution during the analyte determinations. MIP-Gly NPs synthesized using allylamine and squaramide-based monomers appeared most selective to Gly. The Pt-SPEs modified with MIP-Gly NPs were characterized with differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). Changes in the DPV peak originating from the oxidation of the ferrocenyl moieties in these MIP-Gly NPs served as the analytical signal. The DPV limit of detection and the linear dynamic concentration range for Gly were 3.7 pM and 25 pM–500 pM, respectively. Moreover, the selectivity of the fabricated chemosensors was sufficiently high to determine Gly successfully in spiked river water samples.

Development of Molecularly Imprinted Polymer Nanoparticles with High Affinity and Its Application to Electrochemical Biosensors

In this study, redox-labeled molecularly imprinted polymer nanoparticles (nano-MIPs) were synthesized using a solid-phase approach, templated with biomarkers for diabetes such as human serum albumin (HSA). The purpose of this research was to develop enzyme-/antibody-free electrochemical biosensors with the nano-MIP-coated Au electrodes. The synthesized HSA–nano-MIP (HNM) was analyzed for its chemical composition, size, affinity, and electrochemical property, and was tethered on gold via a coupling chemistry. The results of cyclic voltammetry analysis revealed that electron transfer occurred on the HNM-immobilized Au electrode through physical displacement of the redox centers within HNM, which hopped from one redox center to an adjacent redox center, a process known as bounded diffusion. In this paper, we present a promising approach to developing sensitive and selective electrochemical biosensors for diabetes detection.

Dietary powder and molecular imprinted polymer nanoencapsulated sodium propionate to enhance growth performance, digestive enzymes activity, antioxidant defense, and mucosal immune response in African cichlid (Labidochromis lividus) fingerlings

Abstract This study was conducted to examine the effects of powder sodium propionate (P-SP) and SP- loaded molecular imprinted polymer (MIP) nanoparticles (MIP-SP NPs) on the growth, skin mucosal immune parameters, and digestive and liver enzymes activities of African cichlid (Labidochromis lividus) fingerlings. Fish with an average weight of 500±2 mg were stocked into 12 experimental units and fed on experimental diets prepared by supplementing the basal diet (control) with MIP NPs, P-SP (5 g SP kg−1 of dry diet), and MIP-SP NPs for 8 weeks. The findings demonstrated that growth indices improved in the MIP-SP NPs followed by the P-SP dietary group compared to the control groups (P<0.05). The activity of digestive enzymes of lipase, trypsin, protease, and alkaline phosphatase was higher in the fish fed SP-supplemented diets than in the controls (P<0.05). The protease and lipase activities in the MIP-SP NPs dietary group increased by 29.33% and 48.81% compared to the control, respectively (P<0.05). In addition, the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels of liver tissue decreased in the SP dietary groups, while the catalase (CAT ), superoxide dismutase (SOD), and alkaline phosphatase (ALP) levels increased compared to the control groups (P<0.05). The highest SOD and ALP levels were observed in the fish fed on the MIP-SP NPs-supplemented diet (P<0.05). Furthermore, the skin mucosal immune indices, including alternative hemolytic complement activity (ACH50), lysozyme, and total immunoglobulin (Ig) levels increased in the MIP-SP NPs and P-SP dietary groups compared to the controls (P<0.05). The findings indicated that sodium propionate encapsulated in molecularly imprinted polymer nanoparticles could enhance the efficiency of dietary SP in African cichlid fish.

Synthesis of conducting molecularly imprinted polymer nanoparticles for estriol chemosensing

Herein, electrochemically active molecularly imprinted polypyrrole nanoparticles (PPy NPs) were synthesized with the aid of micelles. Iron(III) p-toluenesulfonate induced polymerization of pyrrole resulting in the imprinting of estriol in growing NPs. Two separate imprinted NPs were prepared with pristine and derivatized pyrrole monomers. Techniques such as scanning electron microscopy (SEM), and transmission electron microscopy (TEM), confirmed the synthesis of imprinted PPy NPs of ∼60 nm in size. NPs prepared with pristine pyrrole showed an imprinting factor of 4.2, while NPs prepared with derivatized pyrrole showed no imprinting. The electrochemical-based transduction allowed for the determination of estriol in the concentration range of 0.5–5 µM and 10–100 µM with the 0.16 µM limit of detection (LOD). Furthermore, the selectivity of the MIP NP-coated electrode toward estriol and its structural analogs, namely β-estradiol and estrone, was satisfactory.

Synthesis of fluorescent Molecularly Imprinted Polymer Nanoparticles Sensing Small Neurotransmitters with High Selectivity Using Immobilized Templates with Regulated Surface Density.

To develop nanosensors to probe neurotransmitters, we synthesized fluorescent-functionalized molecularly imprinted polymeric nanoparticles (fMIP-NPs) using monoamine neurotransmitters (serotonin and dopamine) immobilized on glass beads as templates. The size and fluorescence intensity of the fMIP-NPs synthesized with blended silane couplers increased with the presence of the target but were insensitive to the target analogs (L-tryptophan and L-dopa, respectively). However, when the template is anchored by a pure silane agent, both the fluorescence intensity and particle size of the fMIP-NPs were sensitive to the structural analog of the template. Another fMIP-NP was synthesized in the presence of poly([2-(methacryloyloxy)ethyl] trimethylammonium chloride (METMAC)-co-methacrylamide) grafted onto glass beads as a dummy template for acetylcholine. Acetylcholine increased the diameter and fluorescence intensity of the fMIP-NP, but choline had no effect. When the homopolymer of METMAC was used as a template, the fluorescence intensity and size of the resulting nanoparticles were not responsive to either acetylcholine or choline. The principle of increased fluorescence intensity due to specific interaction with the target substance is probably due to the increased distance between the fluorescent functional groups and decreased self-quenching due to the swelling caused by the specific interaction with the template. The results also indicate that MIP nanoparticles prepared by solid-phase synthesis can be used for targeting small molecules, such as the neurotransmitters addressed in this study, by adjusting the surface density of the template.

A rapid synthesis of molecularly imprinted polymer nanoparticles for the extraction of performance enhancing drugs (PIEDs).

It is becoming increasingly more significant to detect and separate hormones from water sources, with the development of synthetic recognition materials becoming an emerging field. The delicate nature of biological recognition materials such as the antibodies means the generation of robust viable synthetic alternatives has become a necessity. Molecularly imprinted nanoparticles (NanoMIPs) are an exciting class that has shown promise due the generation of high-affinity and specific materials. While nanoMIPs offer high affinity, robustness and reusability, their production can be tricky and laborious. Here we have developed a simple and rapid microwaveable suspension polymerisation technique to produce nanoMIPs for two related classes of drug targets, Selective Androgen Receptor Modulators (SARMs) and steroids. These nanoMIPs were produced using one-pot microwave synthesis with methacrylic acid (MAA) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as a suitable cross-linker, producing particles of an approximate range of 120-140 nm. With the SARMs-based nanoMIPs being able to rebind 94.08 and 94.46% of their target molecules (andarine, and RAD-140, respectively), while the steroidal-based nanoMIPs were able to rebind 96.62 and 96.80% of their target molecules (estradiol and testosterone, respectively). The affinity of nanoMIPs were investigated using Scatchard analysis, with Ka values of 6.60 × 106, 1.51 × 107, 1.04 × 107 and 1.51 × 107 M-1, for the binding of andarine, RAD-140, estradiol and testosterone, respectively. While the non-imprinted control polymer (NIP) shows a decrease in affinity with Ka values of 3.40 × 104, 1.01 × 104, 1.83 × 104, and 4.00 × 104 M-1, respectively. The nanoMIPs also demonstrated good selectivity and specificity of binding the targets from a complex matrix of river water, showing these functional materials offer multiple uses for trace compound analysis and/or sample clean-up.

Synergistically Enhancing Tumor Chemotherapy Using an Aggregation-Induced Emission Photosensitizer on Covalently Conjugated Molecularly Imprinted Polymer Nanoparticles.

Due to the polygenic and heterogeneous nature of the tumorigenesis process, traditional chemotherapy is far from desirable. Fabricating multifunctional nanoplatforms integrating photodynamic effect can synergistically enhance chemotherapy because they can make the cancer cells much sensitive to chemotherapeutics. However, how to assemble different units in nanoplatforms and minimize side effects caused by chemodrugs and photosensitizers (PSs) still needs to be explored. Herein, a nanoplatform CPP/PS-MIP@DOX is developed using a simultaneously covalently conjugated new aggregation-induced emission (AIE) PS and a cell-penetrating peptide (CPP) on the surface of silica-based molecularly imprinted polymer (MIP) nanoparticles, prepared with doxorubicin (DOX) as the template in the water system via a sol-gel technique. CPP/PS-MIP@DOX has good biocompatibility, high DOX-loading ability, promoted cellular uptake, and sustained and pH-sensitive drug release capability. Furthermore, it can efficiently penetrate into tumor tissue, accurately home to, and accumulate at the tumor site. As a result, a better efficacy with lower cytotoxicity is achieved with a smaller dosage of DOX by utilizing either the photodynamic effect or unique characteristics of the MIP. It is the first nanoplatform fabricated by chemically conjugating AIE PSs directly on the surface of the scaffold via the surface-decorated strategy and successfully applied in cancer therapy. This work provides an effective strategy by constructing AIE PS-based cancer nanomedicines with MIPs as scaffolds.

Rapid Conductometric Detection of SARS-CoV-2 Proteins and Its Variants Using Molecularly Imprinted Polymer Nanoparticles.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) biosensors have captured more attention than the conventional methodologies for SARS-CoV-2 detection due to having cost-effective platforms and fast detection. However, these reported SARS-CoV-2 biosensors suffer from drawbacks including issues in detection sensitivity, degradation of biomaterials on the sensor's surface, and incapability to reuse the biosensors. To overcome these shortcomings, molecularly imprinted polymer nanoparticles (nanoMIPs) incorporated conductometric biosensor for highly accurate, rapid, and selective detection of two model SARS-CoV-2 proteins: (i) receptor binding domain (RBD) of the spike (S) glycoprotein and (ii) full length trimeric spike protein are introduced. In addition, these biosensors successfully responded to several other SARS-CoV-2 RBD spike protein variants including Alpha, Beta, Gamma, and Delta. Our conductometric biosensor selectively detects the two model proteins and SARS-CoV-2 RBD spike protein variant samples in real-time with sensitivity to a detection limit of 7 pg mL-1 within 10 min of sample incubation. A battery-free, wireless near-field communication (NFC) interface is incorporated with the biosensor for fast and contactless detection of SARS-CoV-2 variants. The smartphone enabled real-time detection and on-screen rapid result for SARS-CoV-2 variants can curve the outbreak due to its ability to alert the user to infection in real time.

Nanoparticle-induced enhancement of cholinesterase activity in the presence of malathion: A potential nerve agent therapeutic

Organophosphate nerve agents are associated with assassination, terrorism and chemical warfare, but there has been slow progress in developing a broad-spectrum response to poisoning. For some nerve agents the oxime component of the therapy may not be effective, limiting the effectiveness of emergency treatment that is desperately needed. An alternative therapy may be possible based on accelerating enzyme (acetylcholinesterase) catalysis in unaffected adjacent enzymes. Herein we demonstrate a restoration of acetylcholinesterase activity in malathion-inhibited cell membrane preparations by the administration of functional nanoparticles. The molecularly imprinted polymer nanoparticles were designed to bind selectively to designated enzyme epitopes. Enzyme activity of membrane-bound acetylcholinesterase was measured in the presence of the organophosphate malathion and the selected nanoparticles. Enzymatic acceleration of the cholinesterase was observed at 162 ± 17 % the rate of erythrocyte ghosts without bound nanoparticles. This may restore sufficient acetylcholine hydrolysis to mitigate the effects of poisoning, offsetting the acetylcholine accumulation resulting from enzyme inhibition.

Synthesis of Molecularly Imprinted Polymer Nanoparticles for SARS-CoV-2 Virus Detection Using Surface Plasmon Resonance

COVID-19 caused by a SARS-CoV-2 infection was first reported from Wuhan, China, and later recognized as a pandemic on March 11, 2020, by the World Health Organization (WHO). Gold standard nucleic acid and molecular-based testing have largely satisfied the requirements of early diagnosis and management of this infectious disease; however, these techniques are expensive and not readily available for point-of-care (POC) applications. The COVID-19 pandemic of the 21st century has emphasized that medicine is in dire need of advanced, rapid, and cheap diagnostic tools. Herein, we report on molecularly imprinted polymer nanoparticles (MIP-NPs/nanoMIPs) as plastic antibodies for the specific detection of SARS-CoV-2 by employing a surface plasmon resonance (SPR) sensor. High-affinity MIP-NPs directed against SARS-CoV-2 were manufactured using a solid-phase imprinting method. The MIP-NPs were then characterized using dynamic light scattering (DLS) and atomic force microscopy (AFM) prior to their incorporation into a label-free portable SPR device. Detection of SARS-CoV-2 was studied within a range of 104–106 PFU mL−1. The MIP-NPs demonstrated good binding affinity (KD = 0.12 pM) and selectivity toward SARS-CoV-2. The AFM, cyclic voltammetry, and square-wave voltammetry studies revealed the successful stepwise preparation of the sensor. A cross-reactivity test confirmed the specificity of the sensor. For the first time, this study demonstrates the potential of molecular imprinting technology in conjunction with miniaturized SPR devices for the detection of SARS-CoV-2 particles with high-affinity and specificity. Such sensors could help monitor and manage the risks related to virus contamination and infections also beyond the current pandemic.

A new electrocatalytic system based on copper (II) chloride and magnetic molecularly imprinted polymer nanoparticles in 3D printed microfluidic flow cell for enzymeless and Low-Potential cholesterol detection

• A new system for the enzymeless electrocatalytic detection of cholesterol has been proposed. • CuCl 2 in acetonitrile served as an effective low potential electrocatalyst for cholesterol oxidation. • Magnetic molecularly imprinted polymer nanoparticles were synthesized and applied for selective cholesterol isolation. • The developed enzymeless electrocatalytic system was validated by using of model mixtures imitating the blood serum. • Microfluidic flow cell was fabricated by 3D printing to perform cholesterol determination. The cholesterol level in blood is an important biomarker for the early diagnosis of atherosclerosis and cardiovascular disease. This paper reports on a new enzymeless electrocatalytic system for the determination of cholesterol. Copper (II) chloride was first used for the enzymeless electrochemical oxidation of cholesterol at a low electrode potential in acetonitrile, and the mechanism for the oxidation process was proposed. Magnetic molecularly imprinted polymer nanoparticles were then synthesized and applied in a 3D printed microfluidic flow cell for selective cholesterol isolation and subsequent analysis. The proposed enzymeless electrocatalytic system allowed for the detection of cholesterol in model solutions with a detection limit of 4 µM and a degree of extraction of more than 90 %. The sensitivity, stability, and duration of the analysis were not inferior to analogs.