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Impaired expression of GABA transporters (GATs) is closely related to the pathogenesis of among others Parkinson's disease and epilepsy. As such, lipophilic nipecotic acid analogs have been extensively studied as GAT1-addressing drugs and radioligands but suffer from limited brain uptake due to the zwitterionic properties of the nipecotic acid moiety. Bioisosteric replacement of the carboxylic acid group is a promising strategy to improve the brain uptake, though it requires knowledge on the binding of these isosteres to GAT1. To screen nipecotic acid isosteres for their affinity to GAT1 in a time- and cost-effective manner, this research aims to develop a molecular imprinted polymer (MIP) that mimics the natural binding site of GAT1 and can act as an alternative screening tool to the current radiometric and mass spectrometry cellular-based assays. To this end, a nipecotic acid MIP was created using the electropolymerization of ortho-phenylenediamine (oPD) by cyclic voltammetry (CV). The optimization of the generated receptor layer was achieved by varying the scan rate (50-250 mV/s) and number of CV cycles (5-12), yielding an optimized MIP with an average imprinting factor of 2.6, a linear range of 1-1000 nm, and a theoretical LOD of 0.05 nm, as analyzed by electrical impedance spectroscopy (EIS). Selectivity studies facilitated the investigation of major binding interactions between the MIP and the substrate, building an experimental model that compares characteristics of various analogs. Results from this model indicate that the substrate carboxylic acid group plays a more important role in binding than an amine group, after comparing the binding of cyclohexanecarboxylic acid (average IF of 1.7) and piperidine (average IF of 0.46). The research culminates in a discussion regarding the feasibility of the in vitro model, comparing the synthetic system against the biological performance of GAT1. Thus, evaluating if it is possible to generate a synthetic GAT1 mimic, and if so, provide directions for follow-up research.
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Polímeros Molecularmente Impressos , Ácidos Nipecóticos , Polímeros Molecularmente Impressos/química , Ácidos Nipecóticos/química , Ácidos Nipecóticos/metabolismo , Humanos , Proteínas da Membrana Plasmática de Transporte de GABA/metabolismo , Proteínas da Membrana Plasmática de Transporte de GABA/química , Impressão MolecularRESUMO
Ensuring a rapid and accurate identification of harmful bacteria is crucial in various fields including environmental monitoring, food safety, and clinical diagnostics. Conventional detection methods often suffer from limitations such as long analysis time, complexity, and the need for qualified personnel. Therefore, a lot of research effort is devoted to developing technologies with the potential to revolutionize the detection of pathogenic bacteria by offering rapid, sensitive, and user-friendly platforms for point-of-care analysis. In this light, biosensors have gained significant commercial attention in recent years due to their simplicity, portability, and rapid analysis capabilities. The purpose of this review is to identify a trend by analyzing which biosensor technologies have become commercially successful in the field of bacteria detection. Moreover, we highlight the characteristics that a biosensor must possess to finally arrive in the market and therefore in the hands of the end-user, and we present critical examples of the market applications of various technologies. The aim is to investigate the reason why certain technologies have achieved commercial success and extrapolate these trends to the future economic viability of a new subfield in the world of biosensing: the development of biomimetic sensor platforms. Therefore, an overview of recent advances in the field of biomimetic bacteria detection will be presented, after which the challenges that need to be addressed in the coming years to improve market penetration will be critically evaluated. We will zoom into the current shortcomings of biomimetic sensors based on imprinting technology and aptamers and try to come up with a recommendation for further development based on the trends observed from previous commercial success stories in biosensing.
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Excitatory amino acid transporters (EAATs) are important regulators of amino acid transport and in particular glutamate. Recently, more interest has arisen in these transporters in the context of neurodegenerative diseases. This calls for ways to modulate these targets to drive glutamate transport, EAAT2 and EAAT3 in particular. Several inhibitors (competitive and noncompetitive) exist to block glutamate transport; however, activators remain scarce. Recently, GT949 was proposed as a selective activator of EAAT2, as tested in a radioligand uptake assay. In the presented research, we aimed to validate the use of GT949 to activate EAAT2-driven glutamate transport by applying an innovative, impedance-based, whole-cell assay (xCELLigence). A broad range of GT949 concentrations in a variety of cellular environments were tested in this assay. As expected, no activation of EAAT3 could be detected. Yet, surprisingly, no biological activation of GT949 on EAAT2 could be observed in this assay either. To validate whether the impedance-based assay was not suited to pick up increased glutamate uptake or if the compound might not induce activation in this setup, we performed radioligand uptake assays. Two setups were utilized; a novel method compared to previously published research, and in a reproducible fashion copying the methods used in the existing literature. Nonetheless, activation of neither EAAT2 nor EAAT3 could be observed in these assays. Furthermore, no evidence of GT949 binding or stabilization of purified EAAT2 could be observed in a thermal shift assay. To conclude, based on experimental evidence in the present study GT949 requires specific assay conditions, which are difficult to reproduce, and the compound cannot simply be classified as an activator of EAAT2 based on the presented evidence. Hence, further research is required to develop the tools needed to identify new EAAT modulators and use their potential as a therapeutic target.
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Transportador 2 de Aminoácido Excitatório , Ácido Glutâmico , Transportador 2 de Aminoácido Excitatório/metabolismo , Impedância Elétrica , Ácido Glutâmico/metabolismo , Transporte Biológico , Transportador 3 de Aminoácido Excitatório/metabolismoRESUMO
By clearing GABA from the synaptic cleft, GABA transporters (GATs) play an essential role in inhibitory neurotransmission. Consequently, in vivo visualization of GATs can be a valuable diagnostic tool and biomarker for various psychiatric and neurological disorders. Not surprisingly, in recent years several research attempts to develop a radioligand have been conducted, but so far none have led to suitable radioligands that allow imaging of GATs. Here, we provide an overview of the radioligands that were developed with a focus on GAT1, since this is the most abundant transporter and most of the research concerns this GAT subtype. Initially, we focus on the field of GAT1 inhibitors, after which we discuss the development of GAT1 radioligands based on these inhibitors. We hypothesize that the radioligands developed so far have been unsuccessful due to the zwitterionic nature of their nipecotic acid moiety. To overcome this problem, the use of non-classical GAT inhibitors as basis for GAT1 radioligands or the use of carboxylic acid bioisosteres may be considered. As the latter structural modification has already been used in the field of GAT1 inhibitors, this option seems particularly viable and could lead to the development of more successful GAT1 radioligands in the future.
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Pseudomonas aeruginosa is a ubiquitous multi-drug-resistant bacterium, capable of causing serious illnesses and infections. This research focuses on the development of a thermal sensor for the indirect detection of P. aeruginosa infection using molecularly imprinted polymers (MIPs). This was achieved by developing MIPs for the detection of pyocyanin, the main toxin secreted by P. aeruginosa. To this end, phenazine was used as a dummy template, evaluating several polymeric compositions to achieve a selective MIP for pyocyanin recognition. The sensitivity of the synthesized MIPs was investigated by UV-vis analysis, with the best composition having a maximum rebinding capacity of 30 µmol g-1 and an imprinting factor (IF) of 1.59. Subsequently, the MIP particles were immobilized onto planar aluminum chips using an adhesive layer, to perform thermal resistance measurements at clinically relevant concentrations of pyocyanin (1.4-9.8 µM), achieving a limit of detection (LoD) of 0.347 ± 0.027 µM. The selectivity of the sensor was also scrutinized by subjecting the receptor to potential interferents. Furthermore, the rebinding was demonstrated in King's A medium, highlighting the potential of the sensor for the indirect detection of P. aeruginosa in complex fluids. The research culminates in the demonstration of the MIP-based sensor's applicability for clinical diagnosis. To achieve this goal, an experiment was performed in which the sensor was exposed to pyocyanin-spiked saliva samples, achieving a limit of detection of 0.569 ± 0.063 µM and demonstrating that this technology is suitable to detect the presence of the toxin even at the very first stage of its production.
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Impressão Molecular , Polímeros Molecularmente Impressos , Pseudomonas aeruginosa , Piocianina , Técnicas EletroquímicasRESUMO
Accurate and fast on-site detection of harmful microorganisms in food products is a key preventive step to avoid food-borne illness and product recall. In this study, screen-printed electrodes (SPEs) were functionalized via a facile strategy with surface imprinted polymers (SIPs). The SIP-coated SPEs were used in combination with the heat transfer method (HTM) for the real-time detection of Escherichia coli. The sensor was tested in buffer, with a reproducible and sensitive response that attained a limit of detection of 180 CFU/mL. Furthermore, selectivity was assessed by analyzing the sensor's response to C. sakazakii, K. pneumoniae and S. aureus as analogue strains. Finally, the device was successfully used for the detection of E. coli in spiked milk as proof-of-application, requiring no additional sample preparation. These results suggest the proposed thermal biosensor possesses the potential of becoming a tool for routine, on-site monitoring of E. coli in food safety applications.
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Técnicas Biossensoriais , Escherichia coli , Staphylococcus aureus , Eletrodos , Técnicas Biossensoriais/métodos , Laticínios , Limite de DetecçãoRESUMO
Per- and polyfluoroalkyl substances (PFAS) are a class of materials that have been widely used in the industrial production of a wide range of products. After decades of bioaccumulation in the environment, research has demonstrated that these compounds are toxic and potentially carcinogenic. Therefore, it is essential to map the extent of the problem to be able to remediate it properly in the next few decades. Current state-of-the-art detection platforms, however, are lab based and therefore too expensive and time-consuming for routine screening. Traditional biosensor tests based on, e.g., lateral flow assays may struggle with the low regulatory levels of PFAS (ng/mL), the complexity of environmental matrices and the presence of coexisting chemicals. Therefore, a lot of research effort has been directed towards the development of biomimetic receptors and their implementation into handheld, low-cost sensors. Numerous research groups have developed PFAS sensors based on molecularly imprinted polymers (MIPs), metal-organic frameworks (MOFs) or aptamers. In order to transform these research efforts into tangible devices and implement them into environmental applications, it is necessary to provide an overview of these research efforts. This review aims to provide this overview and critically compare several technologies to each other to provide a recommendation for the direction of future research efforts focused on the development of the next generation of biomimetic PFAS sensors.
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Biomimética , Fluorocarbonos , Humanos , Carcinogênese , Carcinógenos , IndústriasRESUMO
In recent years, melamine-sensing technologies have increasingly gained attention, mainly due to the misuse of the molecule as an adulterant in milk and other foods. Molecularly imprinted polymers (MIPs) are ideal candidates for the recognition of melamine in real-life samples. The prepared MIP particles were incorporated into a thermally conductive layer via micro-contact deposition and its response towards melamine was analyzed using the heat-transfer method (HTM). The sensor displayed an excellent selectivity when analyzing the thermal response to other chemicals commonly found in foods, and its applicability in food safety was demonstrated after evaluation in untreated milk samples, demonstrating a limit of detection of 6.02 µM. As the EU/US melamine legal limit in milk of 2.5 mg/kg falls within the linear range of the sensor, it can offer an innovative solution for routine screening of milk samples in order to detect adulteration with melamine. The results shown in this work thus demonstrate the great potential of a low-cost thermal platform for the detection of food adulteration in complex matrices.
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This work presents an imprinted polymer-based thermal biomimetic sensor for the detection of Escherichia coli. A novel and facile bacteria imprinting protocol for polydimethylsiloxane (PDMS) films was investigated, and these receptor layers were functionalized with graphene oxide (GO) in order to improve the overall sensitivity of the sensor. Upon the recognition and binding of the target to the densely imprinted polymers, a concentration-dependent measurable change in temperature was observed. The limit of detection attained for the sensor employing PDMS-GO imprints was 80 ± 10 CFU/mL, a full order lower than neat PDMS imprints (670 ± 140 CFU/mL), illustrating the beneficial effect of the dopant on the thermo-dynamical properties of the interfacial layer. A parallel benchmarking of the thermal sensor with a commercial impedance analyzer was performed in order to prove the possibility of using the developed PDMS-GO receptors with multiple readout platforms. Moreover, S. aureus, C. sakazakii and an additional E. coli strain were employed as analogue species for the assessment of the selectivity of the device. Finally, because of the potential that this biomimetic platform possesses as a low-cost, rapid, and on-site tool for monitoring E. coli contamination in food safety applications, spiked fruit juice was analyzed as a real sample. Reproducible and sensitive results fulfill the limit requirements of the applicable European microbiological regulation.
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Impressão Molecular , Biomimética , Dimetilpolisiloxanos , Escherichia coli , Grafite , Impressão Molecular/métodos , Polímeros/química , Staphylococcus aureusRESUMO
Glucose bio-sensing technologies have received increasing attention in the last few decades, primarily due to the fundamental role that glucose metabolism plays in diseases (e.g., diabetes). Molecularly imprinted polymers (MIPs) could offer an alternative means of analysis to a field that is traditionally dominated by enzyme-based devices, posing superior chemical stability, cost-effectiveness, and ease of fabrication. Their integration into sensing devices as recognition elements has been extensively studied with different readout methods such as quartz-crystal microbalance or impedance spectroscopy. In this work, a dummy imprinting approach is introduced, describing the synthesis and optimization of a MIP toward the sensing of glucose. Integration of this polymer into a thermally conductive receptor layer was achieved by micro-contact deposition. In essence, the MIP particles are pressed into a polyvinyl chloride adhesive layer using a polydimethylsiloxane stamp. The prepared layer is then evaluated with the so-called heat-transfer method, allowing the determination of the specificity and the sensitivity of the receptor layer. Furthermore, the selectivity was assessed by analyzing the thermal response after infusion with increasing concentrations of different saccharide analogues in phosphate-buffered saline (PBS). The obtained results show a linear range of the sensor of 0.0194-0.3300 mM for the detection of glucose in PBS. Finally, a potential application of the sensor was demonstrated by exposing the receptor layer to increasing concentrations of glucose in human urine samples, demonstrating a linear range of 0.0444-0.3300 mM. The results obtained in this paper highlight the applicability of the sensor both in terms of non-invasive glucose monitoring and for the analysis of food samples.
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Impressão Molecular , Glicemia , Automonitorização da Glicemia , Glucose , Humanos , Polímeros Molecularmente ImpressosRESUMO
We demonstrate a novel way of creating three-dimensional microfluidic channels capable of following complex topographies. To this end, substrates with open channels and different geometries were 3D-printed, and the open channels were consecutively closed with a thermoplastic using a low-resolution vacuum-forming approach. This process allows the sealing of channels that are located on the surface of complex multiplanar topographies, as the thermoplastic aligns with the surface-shape (the macrostructure) of the substrate, while the microchannels remain mostly free of thermoplastic as their small channel size resists thermoplastic inflow. This new process was analyzed for its capability to consistently close different substrate geometries, which showed reliable sealing of angles >90°. Furthermore, the thermoplastic intrusion into channels of different widths was quantified, showing a linear effect of channel width and percentage of thermoplastic intrusion; ranging from 43.76% for large channels with 2 mm width to only 5.33% for channels with 500 µm channel width. The challenging sealing of substrate 'valleys', which are created when two large protrusions are adjacent to each other, was investigated and the correlation between protrusion distance and height is shown. Lastly, we present three application examples: a serpentine mixer with channels spun around a cuboid, increasing the usable surface area; a cuvette-inspired flow cell for a 2-MXP biosensor based on molecular imprinted polymers, fitting inside a standard UV/Vis-Spectrophotometer; and an adapter system that can be manufactured by one-sided injection molding and is self-sealed before usage. These examples demonstrate how this novel technology can be used to easily adapt microfluidic circuits for application in biosensor platforms.
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Microfluídica , Tratamento de Ferimentos com Pressão Negativa , Polímeros , Impressão TridimensionalRESUMO
The scope of the presented research orientates itself towards the development of a Molecularly Imprinted Polymer (MIP)-based dye displacement assay for the colorimetric detection of the antibiotic amoxicillin in aqueous medium. With this in mind, the initial development of an MIP capable of such a task sets focus on monolithic bulk polymerization to assess monomer/crosslinker combinations that have potential towards the binding of amoxicillin. The best performing composition (based on specificity and binding capacity) is utilized in the synthesis of MIP particles by emulsion polymerization, yielding particles that prove to be more homogenous in size and morphology compared to that of the crushed monolithic MIP, which is an essential trait when it comes to the accuracy of the resulting assay. The specificity and selectivity of the emulsion MIP proceeds to be highlighted, demonstrating a higher affinity towards amoxicillin compared to other compounds of the aminopenicillin class (ampicillin and cloxacillin). Conversion of the polymeric receptor is then undertaken, identifying a suitable dye for the displacement assay by means of binding experiments with malachite green, crystal violet, and mordant orange. Once identified, the optimal dye is then loaded onto the synthetic receptor, and the displaceability of the dye deduced by means of a dose response experiment. Alongside the sensitivity, the selectivity of the assay is scrutinized against cloxacillin and ampicillin. Yielding a dye displacement assay that can be used (semi-)quantitatively in a rapid manner.
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Foodborne illnesses represent high costs worldwide in terms of medical care and productivity. To ensure safety along the food chain, technologies that help to monitor and improve food preservation have emerged in a multidisciplinary context. These technologies focus on the detection and/or removal of either biological (e.g., bacteria, virus, etc.) or chemical (e.g., drugs and pesticides) safety hazards. Imprinted polymers are synthetic receptors able of recognizing both chemical and biological contaminants. While numerous reviews have focused on the use of these robust materials in extraction and separation applications, little bibliography summarizes the research that has been performed on their coupling to sensing platforms for food safety. The aim of this work is therefore to fill this gap and highlight the multidisciplinary aspects involved in the application of imprinting technology in the whole value chain ranging from IP preparation to integrated sensor systems for the specific recognition and quantification of chemical and microbiological contaminants in food samples.
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Impressão Molecular , Polímeros , Inocuidade dos Alimentos , Praguicidas , Receptores ArtificiaisRESUMO
The need to develop interest in STEM (science, technology, engineering, and mathematics) skills in young pupils has driven many educational systems to include STEM as a subject in primary schools. In this work, a science kit aimed at children from 8 to 14 years old is presented as a support platform for an innovative and stimulating approach to STEM learning. The peculiar design of the kit, based on modular components, is aimed to help develop a multitude of skills in the young students, dividing the learning process into two phases. During phase 1 the pupils build the experimental setup and visualize the scientific phenomena, while in phase 2, they are introduced and challenged to understand the principles on which these phenomena are based, guided by a handbook. This approach aims at making the experience more inclusive, stimulating the interest and passion of the pupils for scientific subjects.
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The rapid sensing of drug compounds has traditionally relied on antibodies, enzymes and electrochemical reactions. These technologies can frequently produce false positives/negatives and require specific conditions to operate. Akin to antibodies, molecularly imprinted polymers (MIPs) are a more robust synthetic alternative with the ability to bind a target molecule with an affinity comparable to that of its natural counterparts. With this in mind, the research presented in this article introduces a facile MIP-based dye displacement assay for the detection of (±) amphetamine in urine. The selective nature of MIPs coupled with a displaceable dye enables the resulting low-cost assay to rapidly produce a clear visual confirmation of a target's presence, offering huge commercial potential. The following manuscript characterizes the proposed assay, drawing attention to various facets of the sensor design and optimization. To this end, synthesis of a MIP tailored towards amphetamine is described, scrutinizing the composition and selectivity (ibuprofen, naproxen, 2-methoxphenidine, quetiapine) of the reported synthetic receptor. Dye selection for the development of the displacement assay follows, proceeded by optimization of the displacement process by investigating the time taken and the amount of MIP powder required for optimum displacement. An optimized dose-response curve is then presented, introducing (±) amphetamine hydrochloride (0.01-1 mg mL-1) to the engineered sensor and determining the limit of detection (LoD). The research culminates in the assay being used for the analysis of spiked urine samples (amphetamine, ibuprofen, naproxen, 2-methoxphenidine, quetiapine, bupropion, pheniramine, bromopheniramine) and evaluating its potential as a low-cost, rapid and selective method of analysis.
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Anfetaminas/urina , Corantes/química , Polímeros Molecularmente Impressos , Polímeros/química , Detecção do Abuso de Substâncias/métodos , Urina/química , Anfetamina/urina , Bromofeniramina/urina , Bupropiona/urina , Relação Dose-Resposta a Droga , Técnicas Eletroquímicas , Reações Falso-Positivas , Humanos , Ibuprofeno/urina , Limite de Detecção , Impressão Molecular , Naproxeno/urina , Feniramina/urina , Piperidinas/urina , Pós , Fumarato de Quetiapina/urinaRESUMO
Molecularly imprinted polymers (MIPs) have emerged over the past few decades as interesting synthetic alternatives due to their long-term chemical and physical stability and low-cost synthesis procedure. They have been integrated into many sensing platforms and assay formats for the detection of various targets, ranging from small molecules to macromolecular entities such as pathogens and whole cells. Despite the advantages MIPs have over natural receptors in terms of commercialization, the striking success stories of biosensor applications such as the glucose meter or the self-test for pregnancy have not been matched by MIP-based sensor or detection kits yet. In this review, we zoom in on the commercial potential of MIP technology and aim to summarize the latest developments in their commercialization and integration into sensors and assays with high commercial potential. We will also analyze which bottlenecks are inflicting with commercialization and how recent advances in commercial MIP synthesis could overcome these obstacles in order for MIPs to truly achieve their commercial potential in the near future.
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Point of care (PoC) diagnostics are at the focus of government initiatives, NGOs and fundamental research alike. In high-income countries, the hope is to streamline the diagnostic procedure, minimize costs and make healthcare processes more efficient and faster, which, in some cases, can be more a matter of convenience than necessity. However, in resource-limited settings such as low-income countries, PoC-diagnostics might be the only viable route, when the next laboratory is hours away. Therefore, it is especially important to focus research into novel diagnostics for these countries in order to alleviate suffering due to infectious disease. In this review, the current research describing the use of PoC diagnostics in resource-limited settings and the potential bottlenecks along the value chain that prevent their widespread application is summarized. To this end, we will look at literature that investigates different parts of the value chain, such as fundamental research and market economics, as well as actual use at healthcare providers. We aim to create an integrated picture of potential PoC barriers, from the first start of research at universities to patient treatment in the field. Results from the literature will be discussed with the aim to bring all important steps and aspects together in order to illustrate how effectively PoC is being used in low-income countries. In addition, we discuss what is needed to improve the situation further, in order to use this technology to its fullest advantage and avoid "leaks in the pipeline", when a promising device fails to take the next step of the valorization pathway and is abandoned.
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Sistemas Automatizados de Assistência Junto ao Leito , Doenças Transmissíveis , Recursos em Saúde , Humanos , Monitorização Fisiológica , Testes Imediatos , PobrezaRESUMO
A charge-transfer (CT) interaction between 1,3,5-trinitro-2,4-dimethylbenzene (TNX) and anionic phosphate is evaluated, yielding a high band electronic transfer interaction that can be observed as a distinct color change when phosphate is present in solution. The induced interaction was studied using 1H NMR, UV-visible, and Fourier transform infrared spectroscopies. The stoichiometric determination of the interaction was divined by means of continuous variation, applying the Schaeppi-Treadwell method to calculate the binding constant (k). Furthermore, the effect of the polarity of solvents toward the generation of the CT interaction was examined, with multiple solvents considered. Complex deconstruction studies were undertaken, examining the effects of water on complex destruction and understanding the volumes needed to hinder the CT interaction potency. Specificity and selectivity of the CT interaction were also studied against other biologically relevant species (CH3CH2OH, Na+, K+, Ca2+, Cl-, HCO3 -, F-, CH3COO-, and SO4 2-), assessing the capabilities of the assay to differentiate anionic species and counter cations that could act as interferences. The role of TNX concentration in CT formation was also analyzed, aiming to optimize the phosphate-sensing assay and improve its limit of detection. The sensing platform was subsequently used to study phosphate concentrations in urine samples to further understand its potential application in biomedical research. To validate the developed technique, urine samples were analyzed for their phosphate content with both the developed sensor and a validated vanadate-molybdate reagent. The results indicate that the sensing method is capable of accurately reporting elevated phosphate levels in urine samples in a rapid and sensitive manner, illustrating that the colorimetric test could be used as a prescreening test for conditions such as hyperphosphatemia or chronic kidney disease.
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Given the major structural role phosphodiesters play in the organism it is surprising they have not been more widely adopted as a building block in sophisticated biomimetic hydrogels and other biomaterials. The potential benefits are substantial: phosphoester-based materials show excellent compatibility with blood, cells, and a remarkable resistance to protein adsorption that may trigger a foreign-body response. In this work, a novel class of phosphodiester-based ionic hydrogels is presented which are crosslinked via a phosphodiester moiety. The material shows good compatibility with blood, supports the growth and proliferation of tissue and presents opportunities for use as a drug release matrix as shown with fluorescent model compounds. The final gel is produced via base-induced elimination from a phosphotriester precursor, which is made by the free-radical polymerization of a phosphotriester crosslinker. This crosslinker is easily synthesized via multigram one-pot procedures out of common laboratory chemicals. Via the addition of various comonomers the properties of the final gel may be tuned leading to a wide range of novel applications for this exciting class of materials.
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Liberação Controlada de Fármacos , Ésteres/química , Hidrogéis/química , Alicerces Teciduais/química , Animais , Dimetil Sulfóxido/química , Corantes Fluorescentes/química , Corantes Fluorescentes/metabolismo , Liofilização , Espectroscopia de Ressonância Magnética , Teste de Materiais , Miócitos de Músculo Liso/citologia , SuínosRESUMO
Vitamin K was originally discovered as a cofactor required to activate clotting factors and has recently been shown to play a key role in the regulation of soft tissue calcification. This property of vitamin K has led to an increased interest in novel methods for accurate vitamin K detection. Molecularly Imprinted Polymers (MIPs) could offer a solution, as they have been used as synthetic receptors in a large variety of biomimetic sensors for the detection of similar molecules over the past few decades, because of their robust nature and remarkable selectivity. In this article, the authors introduce a novel imprinting approach to create a MIP that is able to selectively rebind vitamin K1. As the native structure of the vitamin does not allow for imprinting, an alternative imprinting strategy was developed, using the synthetic compound menadione (vitamin K3) as a template. Target rebinding was analyzed by means of UV-visible (UV-VIS) spectroscopy and two custom-made thermal readout techniques. This analysis reveals that the MIP-based sensor reacts to an increasing concentration of both menadione and vitamin K1. The Limit of Detection (LoD) for both compounds was established at 700 nM for the Heat Transfer Method (HTM), while the optimized readout approach, Thermal Wave Transport Analysis (TWTA), displayed an increased sensitivity with a LoD of 200 nM. The sensor seems to react to a lesser extent to Vitamin E, the analogue under study. To further demonstrate its potential application in biochemical research, the sensor was used to measure the absorption of vitamin K in blood serum after taking vitamin K supplements. By employing a gradual enrichment strategy, the sensor was able to detect the difference between baseline and peak absorption samples and was able to quantify the vitamin K concentration in good agreement with a validation experiment using High-Performance Liquid Chromatography (HPLC). In this way, the authors provide a first proof of principle for a low-cost, straightforward, and label-free vitamin K sensor.