Preparation of magnetic glycoprotein‐imprinted nanoparticles with dendritic polyethyleneimine as a monomer for the specific recognition of ovalbumin from egg white

Glycoproteins are crucial in massive physiological events and clinical application. It is necessary to prepare new materials to isolate the specific glycoprotein. New and simple core–shell molecularly imprinted polymers were prepared by surface imprinting. The polymers are synthesized with magnetic nanoparticles as the core, water‐soluble dendritic polyethyleneimine as the monomer and the ovalbumin as the template. The prepared imprinted polymers showed thin imprinted shell, biocompatibility and superparamagnetic properties. The resultant materials exhibited fast kinetics, high adsorption capacity, perfect selectivity and reusability. More important, they can absorb the template glycoprotein from the neutral solution and successfully be applied to recognize the ovalbumin from egg white, which means that they can provide an alternate method to isolate glycoprotein from bodily fluids.     

Selective adsorption of protein by a high‐efficiency Cu2+‐cooperated magnetic imprinted nanomaterial

We report a core–shell magnetic molecularly imprinted polymer with high affinity through a facile sol–gel method for the selective adsorption of bovine hemoglobin from real bovine blood. Copper ions grafted on the surface of the matrix could immobilize template protein through chelation, which greatly enhances the orderliness of imprinted cavities and affinity of polymers. The obtained products exhibit a desired level of magnetic susceptibility, resulting in the highly efficient adsorption process. The results of adsorption experiments show that the saturation adsorption capacity of imprinted products could reach 116.3 mg/g within 30 min. Meanwhile, the specific binding experiment demonstrates the high selectivity of polymers for bovine hemoglobin. Furthermore, satisfactory reusability is demonstrated by ten adsorption–desorption cycles with no obvious deterioration in binding capacity. Electrophoretic analysis suggests the polymer could be used successfully in separation and enrichment of bovine hemoglobin from the bovine blood sample, which exhibits potential application in pretreatment of proteomics.     

Synthesis of magnetic dual‐template molecularly imprinted nanoparticles for the specific removal of two high‐abundance proteins simultaneously in blood plasma

Novel core–shell dual‐template molecularly imprinted superparamagnetic nanoparticles were synthesized using bovine hemoglobin and bovine serum albumin as the templates for the efficient depletion of these two high‐abundance proteins from blood plasma for the first time. The preparation process combined surface imprinting technique and a two‐step immobilized template strategy. The obtained polymers were fully characterized by transmission electron microscopy, X‐ray diffraction, Fourier transform infrared spectroscopy, and vibrating sample magnetometry. The results showed that the as‐synthesized nanomaterials possessed homogeneous and thin imprinted shells with a thickness of about 5 nm, stable crystalline phase, and superparamagnetism. The binding performance of the imprinted polymers was investigated through a series of adsorption experiments, which indicated that the products had satisfactory recognition ability for bovine hemoglobin and bovine serum albumin. The resultant nanoparticles were also successfully applied to simultaneously selective removal of two proteins from a real bovine blood sample.     

Surface molecularly imprinted magnetic microspheres for the recognition of albumin

A new approach, combining metal coordination with the molecular imprinting technique, was developed to prepare affinity materials. Magnetic poly(glycidyl methacrylate) microspheres in monosize form were used for specific recognition toward the target protein. The magnetic poly(glycidyl methacrylate) microspheres were prepared by dispersion polymerization in the presence of magnetite nanopowder. Surface imprinted magnetic poly(glycidyl methacrylate) microspheres based on metal coordination were prepared and used for the selective recognition of human serum albumin. Iminodiacetic acid was used as the metal coordinating agent and human serum albumin was anchored by Cu²⁺ ions on the surface of magnetic poly(glycidyl methacrylate) microspheres by metal coordination. The magnetic poly(glycidyl methacrylate) microspheres were coated with a polymer formed by condensation of tetraethyl orthosilicate and 3‐aminopropyltrimethoxysilane. The human serum albumin imprinted magnetic poly(glycidyl methacrylate) microspheres were characterized by scanning electron microscopy, attenuated total reflectance Fourier transform infrared spectroscopy and particle size analysis. The maximum adsorption capacity of human serum albumin imprinted magnetic poly(glycidyl methacrylate) microspheres was 37.7 mg/g polymer at pH 6.0. The selectivity experiments of human serum albumin imprinted magnetic poly(glycidyl methacrylate) microspheres prepared with different concentrations in the presence of lysozyme, bovine serum albumin and cytochrome C were performed in order to determine the relative selectivity coefficients.     

The effectively specific recognition of bovine serum albumin imprinted silica nanoparticles by utilizing a macromolecularly functional monomer to stabilize and imprint template

Structural stability of the template is one of the most important considerations during the preparation of protein imprinting technology. To address this limitation, we propose a novel and versatile strategy of utilizing macromolecularly functional monomers to imprint biomacromolecules. Results from circular dichroism and synchronous fluorescence experiments reflect the macromolecularly functional monomers tendency to interact with the protein surface instead of permeating it and destroying the hydrogen bonds that maintain the protein’s structural stability, therefore stabilizing the template protein structure during the preparation of imprinted polymers. The imprinted polymers composed of macromolecularly functional monomers or their equivalent micromolecularly functional monomers over silica nanoparticles were characterized and carried out in batch rebinding test and competitive adsorption experiments. In batch rebinding test, the imprinted particles prepared with macromolecularly functional monomers exhibited an imprinting factor of 5.8 compared to those prepared by micromolecularly functional monomers with the imprinting factor of 3.4. The selective and competitive adsorption experiments also demonstrated the imprinted particles made by macromolecularly functional monomers possessed much better selectivity and specific recognition ability for template protein. Therefore, using macromolecularly functional monomers to imprint may overcome the mutability of biomacromolecule typically observed during the preparation of imprinted polymers, and thus promote the further development of imprinting technology.     

Poly(amino acid)-based thermoresponsive molecularly imprinted magnetic nanoparticles for specific recognition and release of lysozyme

In this study, poly(amino acid)-based thermoresponsive molecularly imprinted magnetic nanoparticles for recognition and release of lysozyme was prepared via surface imprinting method. For constructing the molecularly imprinted polymer (MIP) layer, amino acid-based thermoresponsive monomer (N-methacryloyl-l-alanine methyl ester, MA-L-Ala-OMe) was mainly selected for the functional monomer along with N,N′-methylenebis(acrylamide) as the crosslinker. The resultant magnetic MIP nanoparticles were characterized in detail. Meanwhile, the dynamic light scattering studies and swelling ratios measurements were carried out for demonstrating the thermoresponsive property of the imprinted nanoparticles. The prepared magnetic MIP nanoparticles showed good adsorption capacity and selective recognition properties to lysozyme. Moreover, the fast adsorption process could reach equilibrium within 15 min. Importantly, the capture and release of lysozyme could be easily realized simply by altering the temperature of aqueous solution. Furthermore, the prepared imprinted nanoparticles were applied to separate lysozyme from the real egg white samples. The results proved that the thermoresponsive MIPs based on MA-L-Ala-OMe have great potential for selectively enriching target proteins in real samples.     

Efficient synthesis of molecularly imprinted polymers with bio‐recognition sites for the selective separation of bovine hemoglobin

We developed a facile approach to the construction of bio‐recognition sites in silica nanoparticles for efficient separation of bovine hemoglobin based on amino‐functionalized silica nanoparticles grafting by 3‐aminopropyltriethoxylsilane providing hydrogen bonds with bovine hemoglobin through surface molecularly imprinting technology. The resulting amino‐functionalized silica surface molecularly imprinted polymers were characterized using scanning electron microscope, transmission electronic microscopy, Fourier transform infrared spectroscopy, X‐ray photoelectron spectroscopy, and thermogravimetric analysis. Results showed that the as‐synthesized imprinted polymers exhibited spherical morphology and favorable thermal stability. The binding adsorption experiments showed that the imprinted polymers can reach equilibrium within 1 h. The Langmuir isotherm and pseudo‐second‐order kinetic model fitted the adsorption data well. Meanwhile, the imprinted polymers possessed a maximum binding capacity up to 90.3 mg/g and highly selectivity for the recognition of bovine hemoglobin. Moreover, such high binding capacity and selectivity retained after eight cycles, indicating the good stability and reusability of the imprinted polymers. Finally, successful application in the selective recognition of bovine hemoglobin from a real bovine blood sample indicated that the imprinted polymers displayed great potentials in efficient purification and separation of target proteins.     

Synthesis of thermo‐responsive bovine hemoglobin imprinted nanoparticles by combining ionic liquid immobilization with aqueous precipitation polymerization

Surface molecular imprinting over functionalized nanoparticles has proved to be an effective approach for construction of artificial nanomaterials for protein recognition. Herein, we report a strategy for synthesis of core–shell protein‐imprinted nanoparticles by the functionalization of nano‐cores with ionic liquids followed by aqueous precipitation polymerization to build thermo‐responsive imprinted polymer nano‐shells. The immobilized ionic liquids can form multiple interactions with the protein template. The polymerization process can produce thermo‐reversible physical crosslinks, which are advantageous to enhancing imprinting and facilitating template removal. With bovine hemoglobin as a model template, the imprinted nanoparticles showed temperature‐sensitivity in both dispersion behaviors and rebinding capacities. Compared with the ionic‐liquid‐modified core nanoparticles, the imprinted particles exhibited greatly increased selectivity and two orders of magnitude higher binding affinity for the template protein. The imprinted nanoparticles achieved relatively high imprinting factor up to 5.0 and specific rebinding capacity of 67.7 mg/g, respectively. These nanoparticles also demonstrated rapid rebinding kinetics and good reproducibility after five cycles of adsorption–regeneration. Therefore, the presented approach may be viable for the fabrication of high‐performance protein‐imprinted nanoparticles with temperature sensitivity.     

Selective extraction of bisphenol A from water by one‐monomer molecularly imprinted magnetic nanoparticles

One‐monomer molecularly imprinted magnetic nanoparticles were prepared as adsorbents for selective extraction of bisphenol A from water in this study. A single bi‐functional monomer was adopted for preparation of the molecularly imprinted polymer, avoiding the tedious trial‐and‐error optimizations as traditional strategy. Moreover, bisphenol F was used as the dummy template for bisphenol A to avoid the interference from residual template molecules. These nanoparticles showed not only large adsorption capacity and good selectivity to the bisphenol A but also outstanding magnetic response performance. Furthermore, they were successfully used as magnetic solid‐phase extraction adsorbents of bisphenol A from various water samples, including tap water, river water, and seawater. The developed method was found to be much more efficient, convenient, and economical for selective extraction of bisphenol A compared with the traditional solid‐phase extraction. Separation of these nanoparticles can be easily achieved with an external magnetic field, and the optimized adsorption time was only 15 min. The recoveries of bisphenol A in different water samples ranged from 85.38 to 93.75%, with relative standard deviation lower than 7.47%. These results showed that one‐monomer molecularly imprinted magnetic nanoparticles had the potential to be popular adsorbents for selective extraction of pollutants from water.     

Molecularly imprinted magnetic nanoparticles for the micro solid‐phase extraction of thiabendazole from citrus samples

The preparation of molecularly imprinted core–shell magnetic nanoparticles and their subsequent use in the solid‐phase extraction of thiabendazole from citrus sample extracts is described. Molecularly imprinted core–shell magnetic nanoparticles were prepared by the precipitation copolymerization of the imprinting polymerization mixture on the surface of vinyl‐modified silica magnetic nanoparticles and were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. The obtained molecularly imprinted core–shell magnetic nanoparticles exhibited a high selectivity for thiabendazole and were easily collected and separated by an external magnetic field without additional centrifugation or filtration steps. Under optimum conditions, a magnetic molecularly imprinted solid‐phase extraction method was developed allowing the extraction of thiabendazole from citrus sample extracts and final determination by high‐performance liquid chromatography with fluorescence detection. The detection limit was 0.2 mg/kg, far lower than the maximum residue limit established within the European Union for thiabendazole in citrus samples.     

Preparation and application of magnetic molecularly imprinted nanoparticles for the selective extraction of osthole in Libanotis Buchtomensis herbal extract

In this work, novel magnetic molecularly imprinted polymers were prepared for the selective extraction of osthole from Libanotis Buchtomensis herbal extract. During the synthesis process, double bonds grafted on the surface of Fe₃O₄ nanoparticles could not only drive the temple molecules to locate onto the surface of vinyl‐functionalized magnetic nanoparticles by π–π conjugation, which makes the distribution of binding sites ordered, but also direct the occurrence of imprinting polymerization at the surface of magnetic nanoparticles by the copolymerization of vinyl terminal groups with functional monomers and cross‐linking agent. The characteristics of the resulting polymers were evaluated by transmission electron microscopy, X‐ray diffraction, Fourier‐transform infrared spectroscopy, and vibrating sample magnetometry. Adsorption kinetics, isotherms, selectivity, reproducibility, and reusability were discussed, which suggest that the obtained nanomaterials possess rapid binding kinetics, high adsorption capacity of 17.65 mg/g, and favorable selectivity for the target molecule. Satisfactory reproducibility and reusability were verified as well. Meanwhile, the resultant imprinted nanoparticles were successfully applied to selectively separate osthole from the herbal extract, which show great potential in extracting active ingredients from traditional Chinese medicine.     

Thermo‐sensitive surface molecularly imprinted magnetic microspheres based on bio‐macromolecules and their specific recognition of bovine serum albumin

Bovine serum albumin imprinted magnetic microspheres, with functional monomers of modified chitosan, N‐isopropylacrylamide and sulfobetaine methacrylate, were successfully prepared and characterized in detail. Computational analyses showed that during the preparation process, modified chitosan can effortlessly form multiple non‐covalent bonds with protein molecules. Temperature‐sensitive N‐isopropylacrylamide improves the elution efficiency by abating the mass transfer resistance. Meanwhile, the zwitterionic sulfobetaine methacrylate strongly interacts with H₂O molecules, remarkably reducing the non‐specific adsorption. The specific bovine serum albumin adsorption performances of the prepared imprinted material were then determined. The adsorption amount reached 86.87 mg/g and the imprinting factor was 6.49. These excellent specific adsorption properties are attributed to the synergetic effects of the different monomers. The fabricated imprinted material not only exhibits great prospects as a biosensor or separation material for protein molecules, but also provides a collaborative strategy for preparing multi‐functional imprinted materials.     

Highly selective and efficient imprinted polymers based on carboxyl‐functionalized magnetic nanoparticles for the extraction of gallic acid from pomegranate rind

With the combined surface imprinting technique and immobilized template strategy, molecularly imprinted magnetic nanoparticles were successfully prepared and coupled with high‐performance liquid chromatography to selectively separate and determine gallic acid from the pomegranate rind. On the surface of carboxyl‐functionalized magnetic nanospheres, thin imprinting shells were formed using dopamine as monomer and crosslinker. The characteristics, polymerization conditions, and adsorption performances of the resultant nanomaterials were investigated in detail. In addition of good crystallinity, satisfactory magnetism, and uniform morphology of the obtained polymers, they had rapid binding kinetics, high adsorption capacity, and favorable reusability. In the mixed solution of four hydroxybenzoic acids, the prepared nanomaterials have an excellent selectivity to gallic acid with an imprinting factor of as high as 17.5. Therefore, the polymers have great potentials in specific extraction and enrichment of gallic acid from the complex natural resources.     

Organic–inorganic hybrid silica as supporting matrices for selective recognition of bovine hemoglobin via covalent immobilization

The synthesis of poly‐aminophenylboronic acid (APBA) imprinted hybrid silica‐based polymers for selective recognition of bovine hemoglobin (BHb) was described, where the mesoporous hybrid silica supporting matrices were prepared by a mild sol–gel process with tetraethoxysilane and 3‐aminopropyltriethoxysilane as two precursors. Covalent immobilization of BHb was adopted in order to create homogeneous recognition sites. After removal of the template, the resulting imprinted polymers showed high binding affinity toward BHb and the imprinting factor (α) reached 2.12. The specificity of the BHb recognition was evaluated with competitive experiments, indicating the imprinted polymers have a higher selectivity for the template BHb. The easy preparation protocol and good protein recognition properties made the approach an attractive solution to depletion of high‐abundance protein from bovine blood.     

Preparation of “dummy” l‐phenylalanine molecularly imprinted microspheres by using ionic liquid as a template and functional monomer

In this study, dummy imprinting technology was employed for the preparation of l‐ phenylalanine‐imprinted microspheres. Ionic liquids were utilized as both a “dummy” template and functional monomer, and 4‐vinylpyridine and ethylene glycol dimethacrylate were used as the assistant monomer and cross‐linker, respectively, for preparing a surface‐imprinted polymer on poly(divinylbenzene) microspheres. By the results obtained by theoretical investigation, the interaction between the template and monomer complex was improved as compared with that between the template and the traditional l‐ phenylalanine‐imprinted polymer. The batch experiments indicated that the imprinting factor reached 2.5. Scatchard analysis demonstrated that the obtained “dummy” molecularly imprinted microspheres exhibited an affinity of 77.4 M·10^?4, significantly higher that of a traditional polymer directly prepared by l‐ phenylalanine, which is in agreement with theoretical results. Competitive adsorption experiments also showed that the molecularly imprinted polymer with the dummy template effectively isolated l‐ phenylalanine from l‐ histidine and l‐ tryptophan with separation factors of 5.68 and 2.68, respectively. All these results demonstrated that the polymerizable ionic liquid as the dummy template could enhance the affinity and selectivity of molecularly imprinted polymer, thereby promoting the development of imprinting technology for biomolecules.     

Application of a magnetic molecularly imprinted polymer for the selective extraction and trace detection of lamotrigine in urine and plasma samples

Magnetic molecularly imprinted polymers have been synthesized for the selective preconcentration and trace determination of lamotrigine (LTG) in urine and plasma samples. The magnetic nanoparticles were modified by tetraethyl orthosilicate and 3‐methacryloxypropyl trimethoxysilane before imprinting. The magnetic molecularly imprinted polymers were prepared via surface molecular imprinting technique, using Fe₃O₄ as a magnetic component, LTG as template molecule, methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a cross‐linker, and 2,2′‐azobisisobutyronitrile as a radical initiator in methanol/acetonitrile (50:50, v/v) as the porogen. The obtained sorbent was characterized using scanning electron microscopy, Fourier‐transform infrared spectroscopy, X‐ray diffraction, and thermal analysis. Separation of the sorbent from the sample solution was simply achieved by applying an external magnetic field. Determination of the extracted drug was performed by high‐performance liquid chromatography with UV detection. Under the optimum extraction conditions, the method detection limits were 0.7 μg/L (based on S/N of 3) for urine samples and 0.5 μg/L for plasma samples. A linear dynamic range of 1–1000 μg/L was obtained for LTF in plasma and urine samples. Finally, the applicability of the proposed method was evaluated by extraction and determination of LTG in urine and plasma samples.     

Recovery and separation of erythromycin from industrial wastewater by imprinted magnetic nanoparticles that exploit β‐cyclodextrin as the functional monomer

A type of surface imprinting over magnetic Fe₃O₄ nanoparticles utilizing erythromycin‐A as a template for use in the separation and recovery of erythromycin was developed and investigated. As the intermolecular forces play a key role in the performance of imprinted materials, differential scanning calorimetry, and ¹H NMR spectroscopy was employed to evaluate the interactions between erythromycin and the functional monomer β‐cyclodextrin. To synthesize the surface imprinted polymers, magnetic Fe₃O₄ nanoparticles, the core materials, were modified with a free radical initiator to initialize polymerization in a “grafting from” manner. Then using acryloyl‐modified β‐cyclodextrin as the functional monomer and ethyleneglycol dimethacrylate as the cross‐linker, thin erythromycin‐imprinted films were fabricated by the radical‐induced graft copolymerization of monomers on the surface of the Fe₃O₄ nanoparticles. Selectivity experiments showed that the erythromycin‐A‐imprinted materials had recognition ability toward erythromycin derivatives. Finally, these magnetic molecularly imprinted particles were successfully used for the separation and enrichment of erythromycin from the mother liquor. The recovery, detected by high‐performance liquid chromatography and differential pulse voltammetry, approached 97%. The combination of the specific selectivity of the imprinted material and the magnetic separation provided a powerful tool that is simple, flexible, and selective for the separation and recovery of erythromycin.     

Polydopamine‐coated magnetic molecularly imprinted polymer for the selective solid‐phase extraction of cinnamic acid,ferulic acid and caffeic acid from radix scrophulariae sample

We describe novel cinnamic acid polydopamine‐coated magnetic imprinted polymers for the simultaneous selective extraction of cinnamic acid, ferulic acid and caffeic acid from radix scrophulariae sample. The novel magnetic imprinted polymers were synthesized by surface imprinting polymerization using magnetic multi‐walled carbon nanotubes as the support material, cinnamic acid as the template and dopamine as the functional monomer. The magnetic imprinted polymers were characterized by transmission electron microscopy, scanning electron microscopy, Fourier transform infrared spectroscopy and vibrating sample magnetometry. The results revealed that the magnetic imprinted polymers had outstanding magnetic properties, high adsorption capacity, selectivity and fast kinetic binding toward cinnamic acid, ferulic acid and caffeic acid. Coupled with high‐performance liquid chromatography, the extraction conditions of the magnetic imprinted polymers as a magnetic solid‐phase extraction sorbent were investigated in detail. The proposed imprinted magnetic solid phase extraction procedure has been used for the purification and enrichment of cinnamic acid, ferulic acid and caffeic acid successfully from radix scrophulariae extraction sample with recoveries of 92.4–115.0% for cinnamic acid, 89.4–103.0% for ferulic acid and 86.6–96.0% for caffeic acid.     

Molecularly imprinted polymer on a SiO2‐coated graphene oxide surface for the fast and selective dispersive solid‐phase extraction of Carbamazepine from biological samples

A surface carbamazepine‐imprinted polymer was grafted and synthesized on the SiO₂/graphene oxide surface. Firstly SiO₂ was coated on synthesized graphene oxide sheet using the sol–gel technique. Prior to polymerization, the vinyl group was incorporated on to the surface of SiO₂/graphene oxide to direct selective polymerization on the surface. Methacrylic acid, ethylene glycol dimethacrylate and ethanol were used as monomer, cross‐linker and porogen, respectively. Nonimprinted polymer was also prepared for comparison. The properties of the molecularly imprinted polymer were characterized using field‐emission scanning electron microscopy and Fourier‐transform infrared spectroscopy. The surface molecularly imprinted polymer was utilized as an adsorbent of dispersive solid‐phase extraction for separation and preconcentration of carbamazepine. The effects of the different parameters influencing the extraction efficiency, such as sample pH were investigated and optimized. The specificity of the molecular imprinted polymer over the nonimprinted polymer was examined in absence and presence of competitive drugs. The carbamazepine calibration curve showed linearity in the ranges 0.5–500 μg/L. The limits of detection and quantification under the optimized conditions were 0.1 and 0.3 μg/L, respectively. The within‐day and between‐day relative standard deviations (n = 3) were 3.6 and 4.3%, respectively. Furthermore, the relative recoveries for spiked biological samples were above 85%.