Molecularly Imprinted Polymer Layer Research Articles

Dendrimer-like amino-functionalized hierarchical porous silica nanoparticle: A host material for 2,4-dichlorophenoxyacetic acid imprinting and sensing

In this work, a novel molecularly imprinted electrochemical sensor based on the amino-functionalized silica nanoparticles was built for the sensitive and selective detection of 2,4-dichlorophenoxyacetic acid (2,4-D). The hierarchical porous dendrimer-like silica nanoparticles (HPSNs-NH2) were synthesized by an ethyl ether emulsion method. The selective molecularly imprinted polymers (MIP) was prepared on the HPSNs-NH2 modified electrode via electropolymerization by using 2,4-D as the template and o-phenylenediamine (OPD) as the monomer. The porous structure of HPSNs-NH2 reduced the diffusion limitations of the analytes, enhanced the accessibility and increased the surface area of the sensor, while the MIP layer offered the ability to recognize and quantify target 2,4-D by using ferro/ferricyanide as probes. Several significant experimental parameters on the analytical performance of the MIP/HPSNs-NH2 sensor were explored and optimized. Under the optimized condition, the sensor displayed an appreciable selectivity over structurally related compounds and good sensitivity toward 2,4-D. The linear range of 2,4-D detection was from 1.00 × 10−10 to 2.50 × 10−8M and the detection limit was down to 1.17 × 10–11M according to the 3Sa/b criteria. This method has been applied to detect 2,4-D in bean sprout samples with satisfying results.

Integrating fluorescent molecularly imprinted polymer (MIP) sensor particles with a modular microfluidic platform for nanomolar small-molecule detection directly in aqueous samples

Fluorescent sensory MIP (molecularly imprinted polymer) particles were combined with a droplet-based 3D microfluidic system for the selective determination of a prototype small-molecule analyte of environmental concern, 2,4-dichlorophenoxyacetic acid or 2,4-D, at nanomolar concentration directly in water samples. A tailor-made fluorescent indicator cross-linker was thus designed that translates the binding event directly into an enhanced fluorescence signal. The phenoxazinone-type cross-linker was co-polymerized into a thin MIP layer grafted from the surface of silica microparticles following a RAFT (reversible addition-fragmentation chain transfer) polymerization protocol. While the indicator cross-linker outperformed its corresponding monomer twin, establishment of a phase-transfer protocol was essential to guarantee that the hydrogen bond-mediated signalling mechanism between the urea binding site on the indicator cross-linker and the carboxylate group of the analyte was still operative upon real sample analysis. The latter was achieved by integration of the fluorescent core-shell MIP sensor particles into a modular microfluidic platform that allows for an in-line phase-transfer assay, extracting the analyte from aqueous sample droplets into the organic phase that contains the sensor particles. Real-time fluorescence determination of 2,4-D down to 20nM was realized with the system and applied for the analysis of various surface water samples collected from different parts of the world.

Colorimetric detection of pyrethroid metabolite by using surface molecularly imprinted polymer

A simple colorimetric method was developed for the detection of 3-Phenoxybenzaldehyde (3-PBD), the metabolite of pyrethroid pesticides, based on surface molecularly imprinted polymer (MIP). In this method, MIP layer was coated on the surface of monodispersed silica nanoparticles via sol-gel process with 3-Aminopropyltriethoxysilane (APTES) and phenyltrimethoxysilane (PTES) as monomers. Compare with single functional monomer, APTES and PTES can interact with 3-PBD mainly by hydrogen bonding and π-π staking interaction, which makes the synthetic MIPs show better affinity and absorption capacity towards target than that of signal functional monomer. After eluted from MIP nanoparticles, the 3- PBD resulted in a distinctive color fading by potassium permanganate reduction. Taking advantage of this phenomenon, the 3- PBD could be detected by recording the absorption of potassium permanganate solution with MIPs as the recognition element. The MIPs, synthesized under various conditions, were characterized by fourier transform infrared spectrometry and scanning electro microscopy. Meanwhile, their absorption performance were also investigated. Under the optimum condition, the proposed method exhibited a linear concentration range of 3-PBD from 0.1μgmL−1 to 1μgmL−1 with a lower detection limit of 0.052μgmL−1 (S/N=3). Moreover, this method was further used to detect 3-PBD in real samples form river water, fruit juice, and beverage. Satisfactory recovery was achieved in the range of 90.0∼98.9%, which clearly demonstrating the potential value of this strategy in the detection of total pyrethroid pesticides.

TiO2-based Mitoxantrone Imprinted Poly (Methacrylic acid-co-polycaprolctone diacrylate) Nanoparticles as a Drug Delivery System.

Light delivery in photodynamic therapy is a challenging issue in deep cancer treatment. To solve this problem, photosensitizers are conjugated to X-ray luminescent nanoparticles. When the complexes are stimulated by X-rays during radiotherapy, the nanoparticles generate light and activate the photosensitizers. Core-shell molecularly imprinted polymers (MIPs) were prepared against mitoxantrone (MX) in which TiO2 nanoparticles were applied as a core, diacrylated polycaprolctone as a biodegradable cross-linker and methacrylic acid (MAA) or 4-vinylpyridin (4-VP) as the functional monomer. TiO2 was selected as a scintillator, MX as a photosensitizer and MIP as a drug delivery system in order to evaluate the possibility of using photodynamic therapy (PDT) during radiotherapy in the next studies. Binding properties of polymers and drug release profile were studied and the optimized MIP was characterized by SEM, TEM, EDS, FT-IR and XRD. Also, cytotoxicity and free radical production were also studied in vitro. Data indicated that MAA-based MIP had superior binding properties compared to its non-imprinted polymer (NIP) and higher imprinting factor value than MIP-4VP. Drug release experiments indicated higher MX released amount from MAA-based MIP than the other polymers. MAA-based MIP was selected as an optimized carrier for MX delivery system. According to the results, the size of MX-MIP@TiO2 was reported to be less than 75 nm. The free radical production and cytotoxicity of nanoparticles were also evaluated in vitro. The results of the present work proposed the possibility of applying MIP layer as a drug delivery system around TiO2 nanoparticles.

Molecularly imprinted polymer on graphene surface for selective and sensitive electrochemical sensing imidacloprid

A novel and simple imprinting route based on graphene (GN) was proposed to fabricate an electrochemical sensor for sensitive and selective determination of imidacloprid (IDP) residue. In this route, p-vinylbenzoic acid (VBA) was utilized as functional monomer to immobilize onto GN in N,N-dimethylformamide (DMF) solvent via π-π interaction, directing the polymerization of uniform molecularly imprinted polymers (MIP) layers on the surface of GN sheets. Besides, quantitative control of the synthesis of MIP/GN and the preparation of MIP/GN modified glassy carbon electrode (GCE) were carried out, which were rarely found in other papers. Thus, the proposed sensor possesses highly improved stability and specific recognition ability. The adsorption isotherm study indicated that the MIP layer on GN surface is monolayer and the structure is homogeneous. The peak current of IDP on MIP/GN modified GCE is linear with IDP concentration in the range of 0.5–15μM with a detection limit of 0.10μM. In addition, the novel method was further applied to detect IDP in brown rice samples with satisfactory results.

Label-Free Detection of Small Organic Molecules by Molecularly Imprinted Polymer Functionalized Thermocouples: Toward In Vivo Applications.

Molecularly imprinted polymers (MIPs), synthetic polymeric receptors, have been combined successfully with thermal transducers for the detection of small molecules in recent years. However, up until now they have been combined with planar electrodes which limits their use for in vivo applications. In this work, a new biosensor platform is developed by roll-coating MIP particles onto thermocouples, functionalized with polylactic acid (PLLA). As a first proof-of-principle, MIPs for the neurotransmitter dopamine were incorporated into PLLA-coated thermocouples. The response of the synthetic receptor layer to an increasing concentration of dopamine in buffer was analyzed using a homemade heat-transfer setup. Binding of the template to the MIP layer blocks the heat transport through the thermocouple, leading to less heat loss to the environment and an overall higher temperature in the measuring chamber. The measured temperature increase is correlated to the neurotransmitter concentration, which enables measurement of dopamine levels in the micromolar regime. To demonstrate the general applicability of the proposed biosensor platform, thermocouples were functionalized with similar MIPs for cortisol and serotonin, indicating a similar response and limit-of-detection. As the platform does not require planar electrodes, it can easily be integrated in, e.g., a catheter. In this way, it is an excellent fit for the current niche in the market of therapeutics and diagnostics. Moreover, the use of a biocompatible and disposable PLLA-layer further illustrates its potential for in vivo diagnostics.

A potentiometric non-enzymatic glucose sensor using a molecularly imprinted layer bonded on a conducting polymer

A non-enzymatic potentiometric glucose sensor for the determination of glucose in the micomolar level in saliva was developed based on a molecularly imprinted polymer (MIP) binding on a conducting polymer layer. A MIP containing acrylamide, and aminophenyl boronic acid, as a host molecule to glucose, was immobilized on benzoic acid-functionalized poly(terthiophene) (pTBA) by the amide bond formation onto a gold nanoparticles deposited-screen printed carbon electrode (pTBA/AuNPs/SPCE). Aromatic boronic acid was incorporated into the MIP layer to stably capture glucose and create a potentiometric signal through the changed pKa value of polymer film by the formation of boronate anion-glucose complex with generation of H+ ions by the cis-diol reaction. Reversible binding and extraction of glucose on the sensor surface was observed using a quartz crystal microbalance. Each layer of the sensor probe was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, X-ray photoelectron spectroscopy, and atomic force microscopy. The potentiometric response at the optimized conditions exhibited a wide linear dynamic range of 3.2×10−7 to 1.0×10−3M, with a detection limit of 1.9 (±0.15)×10−7M. The sensor probe revealed an excellent selectivity and sensitivity for glucose compared to other saccharides. In addition, the reliability of the proposed glucose sensor was evaluated in physiological fluid samples of saliva and finger prick blood.

Molecularly Imprinted Filtering Adsorbents for Odor Sensing.

Versatile odor sensors that can discriminate among huge numbers of environmental odorants are desired in many fields, including robotics, environmental monitoring, and food production. However, odor sensors comparable to an animal’s nose have not yet been developed. An animal’s olfactory system recognizes odor clusters with specific molecular properties and uses this combinatorial information in odor discrimination. This suggests that measurement and clustering of odor molecular properties (e.g., polarity, size) using an artificial sensor is a promising approach to odor sensing. Here, adsorbents composed of composite materials with molecular recognition properties were developed for odor sensing. The selectivity of the sensor depends on the adsorbent materials, so specific polymeric materials with particular solubility parameters were chosen to adsorb odorants with various properties. The adsorption properties of the adsorbents could be modified by mixing adsorbent materials. Moreover, a novel molecularly imprinted filtering adsorbent (MIFA), composed of an adsorbent substrate covered with a molecularly imprinted polymer (MIP) layer, was developed to improve the odor molecular recognition ability. The combination of the adsorbent and MIP layer provided a higher specificity toward target molecules. The MIFA thus provides a useful technique for the design and control of adsorbents with adsorption properties specific to particular odor molecules.

Electrochemical sensing platform based on molecularly imprinted polymer decorated N,S co-doped activated graphene for ultrasensitive and selective determination of cyclophosphamide

A novel electrochemical sensor has been facilely fabricated by applying composite elements consisting of nitrogen and sulfur co-doped activated graphene (N,S-AGR) and molecularly imprinted polymer (MIP). N,S-AGR was synthesized by one-pot pyrogenation of a mixture of thiourea, KOH and graphene oxide, which was introduced to improve electron transfer capability and surface area of the electrode, while the electro-polymerized MIP layer afforded simultaneous recognition and quantification of cyclophosphamide (CPA) by utilizing Fe(CN)63−/4− as probe to indicate electrical signals. Under the optimal conditions, a calibration curve of current shift versus concentration of CPA was got in the range of 8×10−12–8×10−7molL−1, and the developed sensor gave a remarkably low detection limit (LOD) of 3.4×10−12mol L−1 (S/N=3). Moreover, the as-prepared sensor illustrated other good merits like stability and selectivity, and played a role in real-time therapeutic drug monitoring after CPA administration in rabbit.

One Binder to Bind Them All.

High quality binders, such as antibodies, are of critical importance for chemical sensing applications. With synthetic alternatives, such as molecularly imprinted polymers (MIPs), less sensor development time and higher stability of the binder can be achieved. In this feature paper, I will discuss the impact of synthetic binders from an industrial perspective and I will challenge the molecular imprinting community on the next step to leapfrog the current status quo of MIPs for (bio)sensing. Equally important, but often neglected as an effective chemical sensor, is a good match of transducer and MIP coating for a respective application. To demonstrate an application-driven development, a biosensing use case with surface-imprinted layers on piezoacoustic sensors is reported. Depending on the electrode pattern for the transducer, the strong mechanical coupling of the analyte with the MIP layer coated device allows the adoption of the sensitivity from cell mass to cell viability with complete reversibility.

Synthesis of adenosine-imprinted microspheres for the recognition of ADP-ribosylated proteins

Core-shell structural adenosine-imprinted microspheres were prepared via a two-step procedure. Polystyrene core particles (CP) were firstly prepared via a reversible addition-fragmentation chain transfer (RAFT) polymerization leaving the iniferter on the surface of the cores, then a molecularly imprinted polymer (MIP) shell was synthesized on the surface of the cores by using acrylamide (AAm) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the cross-linker. The formation and growth of the MIP layer were seen dependent on the initiator (AIBN), AAm and the polymerization time used within the polymerization. SEM/TEM images showed that the dimensions of the cores and shells were 2μM and 44nm, respectively. The MIP microspheres exhibited a fast rebinding rate within 2h and a maximum adsorption capacity of 177μg per gram for adenosine. The adsorption fitted a Langmuir-Freundlich (LF) isotherm model with a KLF value of 41mL/μg and a qm value of 177μg/g for the MIP microspheres. The values were larger than those for a non-molecularly imprinted polymer (NIP) particles (5mL/μg and 88μg/g) indicating a better adsorption ability towards adenosine. The MIP microspheres showed a good selectivity for adenosine with a higher adsorption (683nmol/g) for adenosine than that (91nmol/g, 24nmol/g and 54nmol/g) for guanosine, cytidine and uridine respectively. Further experiment proved that the adenosine-imprinted polymer microspheres also had a good selectivity for ADP-ribosylated proteins that the MIP could extract the ADP-ribosylated proteins from the cell extract samples.

8-hydroxy-2′-deoxyguanosine (8-OHdG) biomarker detection down to picoMolar level on a plastic antibody film

An innovative biosensor assembly relying on a simple and straightforward in-situ construction is presented to monitor urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) down to the pmol/L level. The sensing film of the biosensor consisted of a molecularly imprinted polymer (MIP) layer for 8-OHdG assembled on a gold electrode through electropolymerization of monomer combined with the template.The analytical features of the resulting biosensor were assessed by Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). Some experimental parameters such as the initial concentration of the monomer and the ratio template-monomer were investigated and optimized in order to finely tune the performance of the MIP-based sensor. Under optimal conditions, the developed biosensor was able to rebind 8-OHdG with a linear response against EIS from 0.1 to 100pg/ml 3.5-3500 pM. The interference of coexisting species was tested, also with calibrations on urine samples, and good selectivity towards 8-OHdG was obtained.RAMAN spectroscopy, FTIR and SEM evaluations of the prepared films confirmed the formation of a polyphenol thin-film on the electrode surface. The presence and distribution of the imprinted cavities on the MIP layer was confirmed by confocal microscopy imaging of the film, after a post-treatment with Fluorescein Isothiocyanate (FITC) labeled 8-OHdG antibody.Overall, this label-free biosensor for urinary 8-OHdG detection constitutes a promising low-cost alternative to the conventional immunoassay approaches, due to its simplicity, stability, high sensitivity and selectivity for biological sample assays, opening new doors for other applications.

Localized Surface Plasmon Resonance Gas Sensor Based on Molecularly Imprinted Polymer Coated Au Nano-Island Films: Influence of Nanostructure on Sensing Characteristics

A localized surface plasmon resonance gas sensor based on Au nano-island films coated with molecularly imprinted polymer (MIP) layer was developed to selectively detect terpene vapor emitted from plants. Au nano-islands were deposited on a glass substrate through repeated Au sputtering and annealing. The MIP layer was coated on Au nano-island films by spin-coating a pre-polymerized solution containing methacrylic acid as monomer, ethylene glycol dimethylacrylate as cross-linker, and $\alpha $ -pinene as the template molecule. The influence of nanostructure on the refractive index (RI) sensing characteristics was mainly investigated in this paper. The result demonstrated that the structure of Au nano-island films could be controlled by Au sputtering-annealing cycle. Increase in the sputtering-annealing cycle induced the size increase and the inter-particle distance decrease of Au nano-island films. In addition, spectral red shift, decrease in transmittance, and increase in absorbance were observed under this procedure as well. The typical RI sensing evaluation parameters $\Delta \lambda _{\mathrm {{max}}}$ , $\Delta \text{T}_{\mathrm {{min}}}$ , and $\Delta \text{A}_{\mathrm {{max}}}$ achieved the maximum values: 9.75 nm, and 9%, 0.42 under 3 and 18 Au sputtering-annealing cycles. Au nano-island films under 3 and 18 Au sputtering-annealing cycles were coated with the MIP, and the response of MIP-coated Au nano-island sensor to $\alpha $ -pinene vapor was verified to be fast, reversible, and reproducible.

Surface-imprinted polymer coating l-cysteine-capped ZnS quantum dots for target protein specific recognition

This paper demonstrated a new method for preparing fluorescent molecularly imprinted polymer (MIP) for specific recognition of a target protein. The MIP-based fluorescent receptor was developed by coating MIP layer on the surface of l-cysteine modified Mn2+-doped ZnS quantum dots (QDs) using the surface molecular imprinting process. These MIP-QDs composites demonstrated fast adsorption kinetics, high stability, and good dispersibility in aqueous media. Since the fluorescence quenching of MIP-QDs composites is proportional to the concentration of the lysozyme, the MIP-based fluorescent receptor was successfully applied to the direct fluorescence quantification of lysozyme without further pretreatment. The MIP-based fluorescent receptor exhibited good selectivity and sensitivity for lysozyme detection. The optimum fluorescence intensity of the MIP-QDs was found to be at pH 6.0, and the linear range of lysozyme was from 0.1 to 2.0 μM with the detection limit of 25.2 nM. Moreover, the proposed fluorescent receptor was satisfactorily applied to the determination of lysozyme in real samples. This study provides a new approach for recognizing and detecting of specific proteins in biological samples.

Selective determination of trace bisphenol a using molecularly imprinted silica nanoparticles containing quenchable fluorescent silver nanoclusters

We describe a kind of molecularly imprinted polymer (MIP)-based fluorescent sensor. It was obtained by anchoring a surface-imprinted MIP layer for the model analyte bisphenol A (BPA) onto the surface of fluorescent silver nanoclusters (Ag-NCs). The MIP-coated Ag-NCs were synthesized by the sol-gel technique. The hyphenation of the fluorescence of Ag-NCs with MIP technology expands the use of fluorescent nanomaterials in sensing and recognition. BPA is capable of quenching the fluorescence of the Ag-NCs, and the reduction in fluorescence intensity is related to the concentration of BPA in the 0.2 μg mL−1 to 2 mg L−1 range. The detection limit for BPA is 0.02 μg mL−1, and the recovery of BPA from spiked milk and juice samples ranged from 92.5 to 108.5 %. The MIP-coated Ag-NCs presented here are characterized by their ease of synthesis, selectivity and sensitivity. In our perception, this approach can be extended to numerous other analytes.

Monitoring of low levels of furfural in power transformer oil with a sensor system based on a POF-MIP platform.

In this work an innovative, miniaturized and low cost optical chemical sensor (POF-MIP platform), based on a molecular imprinted polymer (MIP) and surface plasmon resonance in a plastic optical fiber (POF), is presented and preliminarily tested for monitoring of furfural (furan-2-carbaldehyde) in transformer oil. To this end, the optical platform was coupled to an MIP layer, highly selective for furfural. The ability of the developed sensor to directly detect furfural in the insulating oil was investigated. The detection limit of the sensor has been found to be 9 ppb, with a linear response up to about 30 ppb. However there is a sensible response up to 0.15 ppm. Because of the small linearity range, the Hill equation is suggested for the quantification. The sensor has been effectively tested in real oil samples collected from aged electrical equipment removed from service. The assessed concentration of furfural is in good agreement with that evaluated by a high pressure liquid chromatography (HLPC) method, confirming the good selectivity of the proposed sensor.

Versatile SERS sensing based on black silicon.

Black Si (b-Si) with gold or silver metal coating has been shown to be an extremely effective substrate for surface-enhanced Raman scattering (SERS). Here, we demonstrate that it is also a highly versatile SERS platform, as it supports a wide range of surface functionalizations. In particular, we report the use of a molecularly imprinted polymer (MIP) coating and a hydrophobic coating on b-Si to establish two different sensing modalities. First, using a MIP layer on Au-coated b-Si, we show selective sensing of two closely related varieties of tetracycline. Second, a hydrophobic coating was used to concentrate the analyte adsorbed on gold colloidal nanoparticles, thus increasing the sensitivity of the measurement by an order of magnitude. In this experiment, Au nanoparticles and analyte were mixed just before SERS measurements and were concentrated by drop-drying on the super-hydrophobic b-Si. These approaches are promising for SERS measurements that are sensitive to the aging of bare plasmonic metal-coated substrates.

Molecularly imprinted polymers-coated gold nanoclusters for fluorescent detection of bisphenol A

A flexible fluorescent sensing strategy for the recognition and detection of bisphenol A (BPA) has been proposed based on molecularly imprinted polymers (MIPs)-coated gold nanoclusters (AuNCs), by taking advantages of the high selectivity of MIPs and the strong fluorescence property of AuNCs. SiO2@AuNCs were initially prepared by making use of the powerful amido bonds between carboxyl-terminated AuNCs and amino-functionalized SiO2 nanoparticles. Then MIPs-coated AuNCs were formed by anchoring MIP layer on the surface of SiO2@AuNCs via a sol–gel process. In the presence of imprinting template BPA, a Meisenheimer complex could be formed between BPA and the primary amino groups on the surface of the AuNCs, and the photoluminescent energy of AuNCs would be transferred to the complex, and thereby result in the fluorescence quenching of AuNCs. The fluorescence-quenching fractions of the sensor presented a satisfactory linearity with BPA concentrations over the range of 0–13.1μM and the detection limit could reach 0.10μM. Distinguished selectivity was also exhibited to BPA over other possibly competing molecules. Moreover, the sensor was successfully applied to determine BPA in seawater, and the average recoveries of BPA at three spiking levels ranged from 91.3 to 96.2% with relative standard deviations below 4.8%. This AuNCs-MIPs based sensor provided great potentials for recognition and determination of phenolic environmental estrogens in complicated samples.

A molecularly imprinted polymer based a lab-on-paper chemiluminescence device for the detection of dichlorvos

In this work, a new molecularly imprinted polymer (MIP) based lab-on-paper device with chemiluminescence (CL) detection of dichlorvos (DDV) was designed. With the circle-shaped device, the MIP layer with certain depth was synthesized and adsorbed on the paper surface and DDV can be selectively imprinted on it. The adsorption and washing procedures can be achieved well on the paper-based chip. The paper-based device was fabricated by a simple cutting method and many chips can be made at the same time. On the basis of DDV enhancing CL of luminol-H2O2 greatly, the proposed MIP based lab-on-paper CL device showed better selectivity to DDV and it has been applied to the determination of DDV in vegetables in the range of 3.0ng/mL–1.0μg/mL with the detection limit of 0.8ng/mL. This study has made a successful attempt in the development of highly selective and sensitive monitoring of DDV in real samples and will provide a new approach for sensitive and specific assay in environmental monitoring.

Improved gate effect enantioselectivity of phenylalanine-imprinted polymers in water by blending crosslinkers

In this work, the anodic current at an electrode grafted with a molecularly imprinted polymer (MIP) crosslinked via a combination of hydrophobic ethyleneglycol dimethacrylate (EDMA) and hydrophilic methylene bisacrylamide (MBAA) was found to exhibit enantioselective sensitivity to the phenylalanine template in aqueous solution. An MIP-grafted electrode crosslinked with a 2:1 mixture of EDMA and MBAA was observed to respond to the template with the highest enantioselectivity, such that the change in current induced by the imprinted template was more than four times that induced by the enantiomer of the template. The contact angle of a water droplet on an MIP-coated electrode prepared using the optimal crosslinker blending ratio was also sensitive to the template and again exhibited chiral selectivity. The change in the contact angle induced by the template was more than twice as large as that obtained from the template’s enantiomer. Atomic force microscopy showed that the surface of the MIP layer fabricated using a mixture of crosslinkers was rougher than that made with a single crosslinking agent, although there was no apparent correlation between the roughness and the enantioselectivity of the layer. These results indicate that the appropriate combination of crosslinkers enables the chiral-selective gate effect by modulating the flexibility and hydrophilicity of the MIP layer. The approach described herein therefore represents a new means of improving the selectivity of MIPs by blending crosslinkers having different chemical properties.