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Gelatin and N-methacryloyl-(L)-histidine methyl ester [MAH] containing poly(2-hydroxyethyl methacrylate) [PHEMA] based cryogels were prepared to aid in enhancing cell proliferation. In order to evaluate effects of MAH comonomer and gelatin, cryogels were prepared containing different amounts of gelatin and MAH. Accordingly, four different groups of cryogels which were PHEMA, poly(2-hydroxyethyl methacrylate)-gelatin [PHEMA/GEL], poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester) [PHEMAH] and poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester)-gelatin [PHEMAH/GEL] were prepared and a total of eight cryogel groups were obtained depending on the varying gelatin ratios. Swelling tests, scanning electron microscope (SEM), microcomputed tomography (Micro-CT) and Fourier transform infrared spectroscopy- Attenuated Total Reflectance (FTIR-ATR) were performed for characterization studies. FTIR-ATR studies confirmed that gelatin and MAH were incorporated into the cryogel structures. Average pore size of cryogels was in the range of 18−198 μm confirmed by Micro-CT and SEM analysis. Then, cryogels were evaluated for cell proliferation abilities by MTT analysis for 24 h and 48 h for cell viability in L929 mouse fibroblast cells. Cell viability of PHEMAH/0.2GEL cryogels reached 209 %. Interaction and proliferation of fibroblast cells on cryogels demonstrated by SEM examination.

In this work, it was aimed to purify peroxidase from Brassica oleracea roots using newly synthesized cryogel membranes. 2-4-vinylphenylboronic acid (VPBA) containing 2-hydroxyethyl methacrylate (HEMA) [PHEMABA] cryogel membranes were prepared successfully and swelling tests, scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, contact angle measurements, and Brauner-Emmet-Teller (BET) surface area analysis were done for characterization. Effect of experimental conditions on peroxidase binding capacity of PHEMABA cryogel membranes was determined by examining parameters such as pH, temperature and time with binding studies. Maximum peroxidase binding capacity of PHEMABA cryogel membranes was determined as 22.01 mg/g for 2 mg/mL equilibrium peroxidase concentration at pH 8.0. Finally, peroxidase was purified from Brassica oleracea roots with 85.38% yield and 18.38 purification fold. According to obtained results, spongy PHEMABA cryogel membranes show high peroxidase binding capacity and could be suggested as a boronate affinity model system for enzyme purification.

Molecularly imprinted cryogels for protein recognition have received much attention in recent years with the development of supermacroporous polymer systems. Molecularly imprinted cryogels, which o...

In the present study, the enantiomeric forms of hydrophobic amino acids ( l -tryptophan, l -tyrosine, and l -phenylalanine) were separated by molecularly imprinted capillary columns (MICC) via Capillary Electrochromatography (CEC) for the first time. The monomer ratio, crosslinker ratio, template molecule ratio, the porogen ratio and type, polymerization time, and also the effect of temperature were examined to increase the permeability properties of MICC. FTIR, SEM and BET analyses were realized for the characterization of MICC. The effect of the electric field, organic solvent ratio, and pressure were carried out experimentally to determine the optimum conditions. The separation performances of MICC and the non-imprinted capillary columns (NICC) were compared electrochromatographically.

Molecularly imprinted polymer-based surface plasmon resonance sensor prepared using silver nanoparticles was designed for the selective recognition of Penicillin G (PEN-G) antibiotic from both aqueous solution and milk sample. PEN-G imprinted sensors (NpMIPs) SPR sensor was fabricated using poly (2-hydroxyethyl methacrylate-N-methacroyl-(L)-cysteine methyl ester)-silver nanoparticles-N-methacryloyl-L-phenylalanine methyl ester polymer by embedding silver nanoparticles (AgNPs) into the polymeric film structure. In addition, a non-imprinted (NpNIPs) SPR sensor was prepared by utilizing the same polymerization recipe without addition of the PEN-G template molecule to evaluate the imprinting effect. FTIR-ATR spectrophotometer, ellipsometer, contact angle measurements were used for the characterization of NpMIPs SPR sensors. The linear concentration range of 0.01–10 ng/mL PEN-G was studied for kinetic analyses. The augmenting effect of AgNPs used to increase the surface plasmon resonance signal response was examined using polymer-based PEN-G imprinted (MIPs) sensor without the addition of AgNPs. The antibiotic amount present in milk chosen as a real sample was measured by spiking PEN-G into the milk. According to the Scatchard, Langmuir, Freundlich and Langmuir–Freundlich adsorption models, the interaction mechanism was estimated to be compatible with the Langmuir model.

In this study, the molecular imprinting method was used to separate enantiomeric forms of chiral antidepressant drug, R,S-citalopram (R,S-CIT) in aqueous solution by CEC system combining the advantages of capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC). For that, an amino acid based molecularly imprinted monolithic capillary column was designed and used as a stationary phase for selective separation of S-citalopram (S-CIT) for the first time. S-CIT was selectively separated from aqueous solution containing the other enantiomeric form of R-CIT, which is same in size and shape to the template molecule. Morphology of the molecularly imprinted (MIP S-CIT) and non-imprinted (NIP S-CIT) monolithic capillary columns was observed by scanning electron microscopy. Imprinting efficiency of MIP S-CIT monolithic capillary column used for selective S-CIT separation was verified by comparing with NIP S-CIT and calculated imprinting factor (I.F:1.81) proved the high selectivity of the MIP S-CIT for S-CIT. Cavities formed for S-CIT form enabled selective (α = 2.08) separation of the target molecule from the other enantiomeric R-CIT form. Separation was achieved in a short period of 10 minutes, with the electrophoretic mobility of 7.68 × 10-6 m2 /Vs for R,S-CIT at pH 7.0 10 mM PB and 50% ACN ratio. The performance of both MIP S-CIT and NIP S-CIT columns was estimated by repeating the R,S-CIT separations with intra-batch and inter-batch studies for reproducibility of retention times of R.S-CITs. Estimated RSD values that are lower than 2% suggest that the monolithic columns separate R,S-CIT enantiomers without losing separation efficiency. This article is protected by copyright. All rights reserved.

In this study, 17β-estradiol (E2) was performed by surface plasmon resonance (SPR) based on molecularly imprinted technique. In the first, SPR sensor was prepared by the formation of E2 imprinted poly(2-hydroxyetyl methacrylate) / (N-methacryloyl-(L)- leucine methyl esters) (PHEMALM) on the modified gold surface of SPR chip. Also, non-imprinted SPR chip was prepared by the same method at the E2 imprinted PHEMALM (E2-MIPHEMALM-SPR) chip without E2. E2-MIPHEMALM-SPR and non-imprinted (NIPHEMALM-SPR) chips were characterized with Fourier transform infrared spectrophotometry (FTIR), ellipsometry, atomic force microscopy (AFM) and contact angle measurements. After that, SPR chips were completed to SPR system to investigate kinetic properties for E2. The sensing ability of E2-MIPHEMALM-SPR chip was investigated with 20-10000 ng/mL concentrations of 17βestradiol solutions. The E2-MIPHEMALM-SPR chip was showed more selectivity for 17β-estradiol than NIPHEMALM-SPR chip. To show the selectivity of E2-MIPHEMALM-SPR chip competitive adsorption of 17β-estradiol, cholesterol and stigmasterol were investigated. E2-MIPHEMALM-SPR chip was investigated ten times with same concentrations of 17β-estradiol solution to show reuse of the chip. The results showed that the E2-MIPHEMALM-SPR chip has high selectivity for 17β-estradiol.

Immunoaffinity chromatography has a huge interest in the biomedical and biotechnological fields, in particular for one-step isolation, purification and removal of analyte compounds. In this study, uniform-sized microcryogels, a new type of cryogels, were synthesized using 2-hydroxyetyhl methacrylate and epoxy-group-containing monomer, glycidyl methacrylate for purification of a plasma protein, transferrin. Immunoaffinity microcryogels containing anti-Tf antibodies were characterized by Raman spectroscopy, Fourier transform infrared spectroscopy, optical microscopy, scanning electron microscopy, density measurements and swelling tests. Adsorption studies in aqueous media were carried out in order to examine the effects of medium pH, initial concentration of analyte and contact time. It was found that the optimum pH was 6.0 and the maximum adsorption capacity of immunoaffinity microcryogels at this pH value found to be 9.82 mg/g. The KL constant for Langmuir isotherm was calculated as 2.65 mL/mg. The maximum adsorption capacity obtained from experimental studies is also very close to the calculated Langmuir adsorption capacity (11.27 mg/g). Langmuir adsorption isotherms and pseudo-second-order kinetic models are consistent with the adsorption process, which means that the adsorption is single layered and chemically controlled. The purity of the eluted hsTf from plasma was about 84% with yield about 82%. After the tenth use of the same microcryogels, the maximum hsTf adsorption capacity decreased by about 20%. The results indicated that the immunoaffinity microcryogels having anti-Tf antibody ligands could be a safe and cost-friendly method for purification of transferrin.

It was aimed to prepare immunoglobulin G (IgG) imprinted thermosensitive p(NIPA-MAH)/IgG-MIP monolithic cryogel for IgG purification. NIPA and MAH was selected as functional comonomers in order to synthesis of thermosensitive IgG imprinted polymer. NIPA is a thermo-responsive monomer with a LCST of 32 °C. It has a hydrophobic character above LCST and hydrophilic character below LCST. MAH provides selective binding on/in the IgG imprinted cavities with the advantage of its ionizable groups depending on change of pH value. After characterization studies, IgG binding capacity of the p(NIPA-MAH) cryogel from aqueous solutions was examined in different experimental conditions. Maximum IgG binding capacity was 106.1 mg/g, obtained at pH 7.4 and 40 °C. The p(NIPA-MAH)/IgG-MIP cryogel was reusable at least ten times. Selectivity of the p(NIPA-MAH) cryogel for IgG was investigated and suitability of the imprinted cryogel for IgG purification from human plasma was confirmed. In conclusion, the resultant cryogel having increased porosity along with high mechanical and chemical strength could meet requirements; thereby these properties make the p(NIPA-MAH)/IgG-MIP cryogel suitable gel matrix for IgG purification. The p(NIPA-MAH)/IgG-MIP cryogel serving as a thermosensitive molecularly imprinted polymer offers high potential for selective recognition of IgG via imprinting and controlled binding-elution opportunity by simple thermoresponsive regulation.

In the study, purification of ovalbumin was performed by modifying polyamide hollow fiber membranes using immobilized metal affinity chromatography technique. For this purpose, firstly polyethyleneimine (PEI) solutions of different concentrations were attached to hollow fiber membranes. Then, Cu(II), Ni(II) and Zn(II) metal ions were chelated separately to polyethyleneimine attached hollow fiber membranes. Characterization studies of modified hollow fiber membranes were performed with scanning electron microscopy (SEM). Also, the surface area was measured with the Brunner Emmet Teller (BET) method and the porosity was measured with mercury porosimeter. pH, ionic strength, initial ovalbumin concentration, temperature and reusability parameters affecting adsorption capacity were investigated. The maximum ovalbumin adsorption capacities of hollow fiber membranes were found to be 317 mg/g for Cu(II), 169 mg/g for Ni(II) and 101 mg/g for Zn(II), respectively. Desorption ratio of metal ions were calculated as 91.6% for Cu(II), 92.9% for Ni(II) and 91.8% for Zn(II), which are quite high and suitable. When examined in terms of adsorption isotherm models, it was concluded that the Langmuir model is suitable. Purification of ovalbumin from egg white was carried out by fast performance liquid chromatography (FPLC), and the purity of ovalbumin was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) method.

Affinity chromatography is a standard method, which used for protein purification and separation studies due to its specificity and selectivity. There are several affinity chromatography methods, such as dye affinity, immobilized metal chelated affinity, and affinity electrophoresis. Cibacron Blue F3G-A (CBD), as a dye ligand, is one of the most used dyes among dye affinity chromatography. CBD is ideally suited for human serum albumin (HSA) separation and purification in affinity chromatography for several years. However, even though CBD has many purification applications, there is not much research focused on the interaction between CBD and HSA in molecular docking. The interactions between CBD and HSA were simulated via AutoDock molecular docking software in this study. Investigated possibilities resulted in six different conformations on different locations, which light the way to variable connectivity of CBD. Thus, it was determined that the most favorable binding is conformation 5, with its lowest binding energy, which is energetically favorable.

In this study, a simple, fast, sensitive and selective surface plasmon resonance (SPR) sensor was prepared using molecular imprinting method for endotoxin detection. Endotoxin imprinted and non-imprinted poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester) based nanofilms were synthesized on the SPR chip surfaces using ultraviolet polymerization. Endotoxin imprinted and non-imprinted SPR sensors were characterized by using contact angle, atomic force microscopy and ellipsometry. After characterization studies, kinetic studies was carried out in the concentration range of 0.5-100 ng/mL. The limit of detection and quantification were obtained as 0.023 and 0.078 ng/mL, respectively. The response time for the equilibration, adsorption and regeneration was approximately 14 min. The selectivity studies with cholesterol and hemoglobin of endotoxin imprinted SPR sensor were examined. Validation studies were carried out via limulus amebocyte lysate (LAL) in order to demonstrate the applicability of the SPR sensor.

WOS:000612823000013 PubMed: 33383498 Boronic acids are important ligands used to selectively recognize and enrich compounds containing cis-diol groups such as nucleosides. In the present study, organic-inorganic hybrid [POSS-MAH-BPA] monolithic column was prepared for the first time in the literature as a new boronate affinity system for the recognition of nucleosides. The selectivity of the [POSS-MAH-PBA] boronate affinity monolithic column for the recognition of cis-diol containing adenosine nucleoside from its analogue molecule of deoxyadenosine triphosphate, dATP, noncis-diol containing compound was investigated both by UV and HPLC studies. When the relative selectivity coefficients are compared, the [POSS-MAH-PBA] boronate affinity monolithic column is 4.25 times more selective for adenosine than [POSS-MAH] monolithic column. Besides, to determine endogenous nucleosides in biological fluids, which may serve as non-invasive cancer biomarkers, nucleosides were spiked into the urine solutions and passed through the [POSS-MAH-PBA] boronate affinity monolithic column, and the nucleosides were confirmed by HPLC. The adenosine recognition capability of the [POSS-MAH-PBA] boronate affinity monolithic column with an average enrichment factor of 48.9-fold was apparently superior to that of the [POSSMAH] monolithic column. Methacryl Polyhedral Oligomeric Silsesquioxanes (POSS-MA) with nano-sized stable 3-dimensional architectures provided the advantage of being used as an adsorbent for the monolithic structure by providing high surface area, 507.60 m(2)/g, and enabling vinyl groups to function with amino acid-based MAH monomers capable of providing electrons to coordinate PBA. Recovery results of more than 90% for adenosine showed that the [POSS-MAH-PBA] boronate affinity monolithic column could be a promising adsorbent for selective adsorption of cis-diol containing compounds such as nucleosides.

Understanding the fundamentals of natural design, structure, and function has pushed the limits of current knowledge and has enabled us to transfer knowledge from the bench to the market as a product. In particular, biomimicry―one of the crucial strategies in this respect―has allowed researchers to tackle major challenges in the disciplines of engineering, biology, physics, materials science, and medicine. It has an enormous impact on these fields with pivotal applications, which are not limited to the applications of biocompatible tooth implants, programmable drug delivery systems, biocompatible tissue scaffolds, organ-on-a-chip systems, wearable platforms, molecularly imprinted polymers (MIPs), and smart biosensors. Among them, MIPs provide a versatile strategy to imitate the procedure of molecular recognition precisely, creating structural fingerprint replicas of molecules for biorecognition studies. Owing to their affordability, easy-to-fabricate/use features, stability, specificity, and multiplexing capabilities, host-guest recognition systems have largely benefitted from the MIP strategy. This review article is structured with four major points: (i) determining the requirement of biomimetic systems and denoting multiple examples in this manner; (ii) introducing the molecular imprinting method and reviewing recent literature to elaborate the power and impact of MIPs on a variety of scientific and industrial fields; (iii) exemplifying the MIP-integrated systems, i.e., chromatographic systems, lab-on-a-chip systems, and sensor systems; and (iv) closing remarks.

Adenosine is an important molecule in the human body because it participates various biochemical processes, signalling in the physiological processes, and neurological disorders. In the current study, the surface imprinting method was used to prepare adenosine-imprinted magnetic core-shell polyvinylbutyral microbeads. These microbeads were utilized for quantification of adenosine in aqueous solution and control plasma in the range of 1–200 µM. The limit of detection was found to be 1.9 nM, which is quite sensitive compared with to some earlier studies. Fourier transform infrared spectroscopy, scanning electron microscopy, and a Zetasizer (particle size analyzer) were used for characterization of the prepared imprinted microbeads. To determine the efficiency of this method, selectivity experiments were conducted with adenosine-imprinted and non-imprinted magnetic core-shell polyvinylbutyral microbeads and with the competitive nucleosides cytidine, uridine, guanosine, and thymidine. Thermodynamic and kinetic studies were performed to assess adsorption of adenosine onto the adenosine-imprinted magnetic core-shell polyvinylbutyral microbeads from adenosine solution. The efficiency was linked to the specific surface reactivity, polarity and porosity of the imprinted microbeads.

In this study, we have reported a novel fabrication technique for human serum albumin (HSA) imprinted composite bacterial cellulose nanofibers (MIP-cBCNFs) used for the depletion of HSA selectively from artificial blood plasma for proteomic applications. Molecular imprinting was achieved by using metal ion coordination interactions of N?methacryloyl?(L)?histidinemethylester (MAH) monomer and Cu(II) ions. MAH-Cu(II)-HSA complex was polymerized with bacterial cellulose nanofibers (BCNFs) under constant stirring at room temperature. The characterization of the MIP-cBCNFs was carried out by FTIR-ATR, SEM, contact angle measurements and surface area measurements. The adsorption experiments of HSA onto the MIP-BCNFs and NIP-BCNFs from aqueous HSA solutions were investigated in a batch system. The selectivity of the MIP-cBCNFs was investigated by using non-template human transferrin (HTR), and myoglobin (Myo). The relative selectivity coefficients of the MIP-cBCNFs were calculated as 4.73 and 3.02 for HSA/HTR and HSA/Myo molecules, respectively. In addition, the depletion of HSA from artificial human plasma was confirmed by SDS-PAGE and 2-D gel electrophoresis. As a result, it has been shown that metal ion coordination interactions contribute to specific binding of template when preparing MIP-cBCNFs for the depletion of HSA with a high adsorption capacity, significant selectivity and reusability. © 2018

In recent years, monolithic columns have been widely used as efficient stationary phases due to some advantages over particle-based counterparts such as their relative ease of preparation a...

Various changes in the environment can occur as drug delivery systems travel to the targeted area. Therefore, it is an important aspect to consider the effects of environmental changes such as pH and temperature on the release in the development of drug delivery systems. Microcryogels having predefined shape and size were fabricated as drug delivery systems and loaded with diclofenac sodium, a non-steroidal anti-inflammatory drug. Physicochemical, structural, and morphological properties of microcryogels were investigated by swelling, pore volume and porosity tests, infrared spectroscopy, and field emission scanning electron microscopy. Effects of medium pH on drug release were examined at release media with the pH 2.0 as simulated gastric juice, pH 7.4 as the simulated intestinal fluid/intravenous solution, pH 6.0 as a simulated medium during a normal wound healing, and pH 9.0 as a simulated medium for infected wounds. Effects of temperature on drug release were also analyzed at 37 °C as normal body temperature and 42 °C as inflammatory temperature. The obtained results showed that drug release was too low in acidic environment than those observed at higher pH values. Moreover, the release rates were higher at inflammatory temperature compared with the ones at normal body temperature.

The potential of the modified bacterial cellulose (BC) nanofibers was determined bearing metal ion coordination interactions to enhance the protein adsorption and binding capacity. Thus, a household synthesized metal chelating monomer, namely N-methacryloyl-L-histidine methylester (MAH), and a commercial metal chelating monomer, namely 4-vinylimidazole (VIm), were used to complex with metal ions Cu(II) and Ni(II) respectively for the synthesis of the modified BC nanofibers. The modified nanofibers were characterized by FT-IR, SEM and EDX measurements. The protein adsorption tests were carried out using hemoglobin as a model protein and it was determined that the maximum adsorption capacity of hemoglobin onto the modified BC nanofibers was found as 47.40 mg/g. The novel strategy for the preparation of metal chelated nanofibers was developed. © 2017 Elsevier B.V.

WOS:000503065300011 In this study, D,L-ampicillin separation was carried out by ligand exchange-micellar electrokinetic chromatography method using L-Lysine monohydrochloride as a ligand and copper (II) sulfate pentahydrate is a central ion supplier. Isomeric separations were performed using capillary electrophoresis (CE) instrument, in which SDS-L-Lys-Cu+2 micelle complex was used as a pseudostationary phase. The effect of pH, SDS amount, applied electrical field, pressure, organic solvent ratio and ampicillin D,L-ratios were investigated. Fast,inexpensive and sensitive approach for the simultaneous separation of D,L-ampicillin in both aqueous and real antibiotic sample was performed using CE coupled with UV detector. The separation was achieved in a short period of 7 minutes with high-sensitivity andlow-detection limit of 1.25 mu M by the developed SDS-L-Lys-Cu+2 micelle-chiral selector complexes without using any extra process such as imprinting or spacer arms for the immobilization of the ligands.

Concanavalin A is a representative of the plant protein group known as lectins. Many lectin proteins have useful characteristics for studies on cell division and cell surfaces. In this study, a new adsorbent for the specific separation of Concanavalin A was prepared by applying a silica particle surface imprinting method. First, silica particles were activated via acidic treatment, and then, 3-methacryloyloxypropyl trimethoxysilane (MPTMS) was used for modification. For the preparation of Concanavalin A surface-imprinted silica particles (Con A-MISPs), N-methacryloyl- l -histidine methyl ester (MAH) was used as a functional monomer. The silica particles were characterized using a Zetasizer, scanning electron microscopy equipment (SEM), and Fourier transform infrared spectroscopy (FTIR). The effects of parameters such as the pH, initial concentration of Concanavalin A, and temperature on the adsorption of Concanavalin A were determined. The maximum Concanavalin A adsorption onto Con A-MISPs was observed to be 305.2 mg/g at a pH of 6. The reusability of the Con A-MISPs was approximately 93.5%. The non-imprinted silica particles (NISPs) were prepared in the same manner without Concanavalin A to compare the surface imprinting factor. Selective binding studies were carried out with lysozyme and hemoglobin molecules. The selectivity of the Con A-MISPs was also investigated by isolating Concanavalin A from Canavalia ensiformis. The purity of the Concanavalin A was shown by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE).

Cryogels have been demonstrated to be efficient when applied for protein isolation. Owing to their macroporous structure, cryogels can also be used for treating particle-containing material, e.g. cell homogenates. Another challenging development in protein purification technology is the use of molecularly imprinted polymers (MIPs). These MIPs are robust and can be used repeatedly. The paper presents a new technology that combine the formation of cryogel beads concomitantly with making imprints of a protein. Protein A was chosen as the print molecule which was also be the target in the purification step. The present paper describes a new method to produce protein-imprinted cryogel beads. The protein-imprinted material was characterized and the separation properties were evaluated with regard to both the target protein and whole cells with target protein exposed on the cell surface. The maximum protein A adsorption was 18.1 mg/g of wet cryogel beads. The selectivity coefficient of protein A-imprinted cryogel beads for protein A was 5.44 and 12.56 times greater than for the Fc fragment of IgG and protein G, respectively.

Sensors have been extensively used owing to multiple advantages, including exceptional sensing performance, user-friendly operation, fast response, high sensitivity and specificity, portability, and real-time analysis. In recent years, efforts in sensor realm have expanded promptly, and it has already presented a broad range of applications in the fields of medical, pharmaceutical and environmental applications, food safety, and homeland security. In particular, molecularly imprinted polymer based sensors have created a fascinating horizon for surface modification techniques by forming specific recognition cavities for template molecules in the polymeric matrix. This method ensures a broad range of versatility to imprint a variety of biomolecules with different size, three dimensional structure, physical and chemical features. In contrast to complex and time-consuming laboratory surface modification methods, molecular imprinting offers a rapid, sensitive, inexpensive, easy-to-use, and highly selective approaches for sensing, and especially for the applications of diagnosis, screening, and theranostics. Due to its physical and chemical robustness, high stability, low-cost, and reusability features, molecularly imprinted polymer based sensors have become very attractive modalities for such applications with a sensitivity of minute structural changes in the structure of biomolecules. This review aims at discussing the principle of molecular imprinting method, the integration of molecularly imprinted polymers with sensing tools, the recent advances and strategies in molecular imprinting methodologies, their applications in medical, and future outlook on this concept.

Synthesis of melamine (ME) imprinted polymeric sorbent for use as solid phase extraction adsorbent was targeted and the benefits of the synthesis in the monolith format were combined with imprinting technology advantages. Melamine as template and 2-vinylimidazole-Cu 2 + as complexing agent were used as the first time for pre-complex formation, followed by the preparation of monolith by bulk polymerization method in the presence of ethyleneglycol dimethacrylate as crosslinker, azobisisobutyronitrile as initiator, and ethyl alcohol as solvent for a duration of 4 h at 80 °C. 1.0 M NaOH solution was used as a desorption agent to remove template. Non-imprinted monolith (NIP) were synthesized as a control without template in same receipt. The monolith samples were characterized by Fourier Transformed Infrared Spectroscopy (FTIR), Scanning Electron Microscope (SEM), elementary analysis, and Branauer- Emmett- Teller (BET) studies. Melamine adsorption from aqueous solution was studied in continuous mode, and some parameters such as pH, temperature, concentration and flow-rate were optimized for melamine adsorption. The maximum adsorption capacities were determined as 147.9 mg/g for imprinted polymer (MIP), 82.0 mg/g for non-imprinted polymer (NIP) and 4.0 mg/g for polymer without pre-complex (E) at 25 °C, and pH 5.0 with 500 ppm melamine initial concentration for 1.0 mL/min flow-rate. In addition to interaction characterized by metal coordination bond formation, non- covalent interactions were responsible for melamine adsorption. No significant change in adsorption performance was recorded after 10 consecutive adsorption-desorption cycles. Kinetic analysis indicated compliance with pseudo-second order kinetics. The Freundlich isotherm was found suitable for description of the process. Cyanuric acid (CY), aniline (AN), and phenol (PH) were used as competing agents in selectivity studies, and relative selectivity coefficients were calculated (k' CY = 4.45; k' AN = 7.08; k' PH = 3.98). The performance of melamine imprinted polymeric monolith (MIP) as solid-phase extraction adsorbent for extraction of melamine from milk samples was investigated and the recovery of melamine in spiked milk samples was 97.6% with the relative standard deviation (RSD) (8.1%).

The publications in macro-molecularly imprinted polymers have increased drastically in recent years with the development of water-based polymer systems. The macroporous structure of cryogels has allowed the use of these materials within different applications, particularly in affinity purification and molecular imprinting based methods. Due to their high selectivity, specificity, efficient mass transfer and good reproducibility, molecularly imprinted cryogels (MICs) have become attractive for researchers in the separation and purification of proteins. In this review, the recent developments in affinity based cryogels and molecularly imprinted cryogels in protein purification are reviewed comprehensively.

Molecular imprinting technology (MIT) presents self-organization of functional monomers around a template, leading formation of specific recognition sites for the template with high selectivity. In this study, cryogels in microscale were fabricated as novel plastic antibodies via MIT for transferrin purification. After chemical and physical characterization studies, optimal hsTf adsorption conditions for the plastic antibodies were then determined by investigating parameters including medium pH and temperature, initial analyte concentration and salt concentration. Selectivity studies were performed by considering both size and isoelectric points of the proteins. Compared with the non-imprinted P(HEMATrp) microcryogels, the relative selectivity coefficients of the plastic antibodies for hsTf/MYB and hsTf/HSA were found to be 2.24 and 2.10 times higher, respectively. The findings indicated that the molecularly imprinted microcryogels could be selective, reusable and cost-friendly potential plastic antibodies for purification of transferrin.

Hemoglobin is an iron carrying protein in erythrocytes and also an essential element to transfer oxygen from the lungs to the tissues. Abnormalities in hemoglobin concentration are closely correlated with health status and many diseases, including thalassemia, anemia, leukemia, heart disease, and excessive loss of blood. Particularly in resource-constrained settings existing blood analyzers are not readily applicable due to the need for high-level instrumentation and skilled personnel, thereby inexpensive, easy-to-use, and reliable detection methods are needed. Herein, a molecular fingerprints of hemoglobin on a nanofilm chip was obtained for real-time, sensitive, and selective hemoglobin detection using a surface plasmon resonance system. Briefly, through the photopolymerization technique, a template (hemoglobin) was imprinted on a monomeric (acrylamide) nanofilm on-chip using a cross-linker (methylenebisacrylamide) and an initiator-activator pair (ammonium persulfate-tetramethylethylenediamine). The molecularly imprinted nanofilm on-chip was characterized by atomic force microscopy and ellipsometry, followed by benchmarking detection performance of hemoglobin concentrations from 0.0005 mg mL&minus, 1 to 1.0 mg mL&minus, 1. Theoretical calculations and real-time detection implied that the molecularly imprinted nanofilm on-chip was able to detect as little as 0.00035 mg mL&minus, 1 of hemoglobin. In addition, the experimental results of hemoglobin detection on the chip well-fitted with the Langmuir adsorption isotherm model with high correlation coefficient (0.99) and association and dissociation coefficients (39.1 mL mg&minus, 1 and 0.03 mg mL&minus, 1) suggesting a monolayer binding characteristic. Assessments on selectivity, reusability and storage stability indicated that the presented chip is an alternative approach to current hemoglobin-targeted assays in low-resource regions, as well as antibody-based detection procedures in the field. In the future, this molecularly imprinted nanofilm on-chip can easily be integrated with portable plasmonic detectors, improving its access to these regions, as well as it can be tailored to detect other proteins and biomarkers.

Aquatic and terrestrial environment and human health have been seriously threatened with the release of metal-containing wastewater by the rapid growth in the industry. There are various methods which have been used for removal of ions from the environment, such as membrane filtration, ion exchange, membrane assisted liquid extraction and adsorption. As a sort of special innovation, a polymerization technique, namely molecular imprinting is carried out by specific identification for the target by mixing it with a functional monomer. After the polymerization occurred, the target ion can be removed with suitable methods. At the end of this process, specific cavities, namely binding sites, are able to recognize target ions selectively. However, the selectivity of the molecularly imprinted polymer is variable not only because of the type of ligand but also charge, size coordination number, and geometry of the target ion. In this review, metal ion-imprinted polymeric materials that can be applied for metal ion removal from different sources are discussed and exemplified briefly with different metal ions.

Molecularly imprinted polymers possessing synthetic recognition sites present great potential for various application areas, which require specific recognition of molecules. However various molecularly imprinted polymers (MIPs) reported previously have low affinity capacity and low binding kinetics due to weak interactions between template molecule and functional monomer in aqueous media and also high mass transfer resistance. The synthesis of MIPs onto nanomaterial surface via metal ion coordination interactions serves as an effective solution. In this work, we prepared thin molecularly imprinted polymer layer on bacterial cellulose nanofiber surface using Cu2+ chelating monomer, N-Methacryloyl-(L)-histidinemethylester (MAH) as a functional monomer orienting cytochrome c molecules. The conditions of polymerization and adsorption were optimized to obtain best affinity performance. Due to unique properties of bacterial cellulose nanofibers such as high surface-to-volume ratio and macro-porosity, the resultant cytochrome c imprinted nanofibers (Cyt c-MIP NFs) reducing diffusional resistances could achieve high recognition capacity and excellent binding kinetics. Cyt c-MIP NFs were also successfully applied to capture template proteins from real sample. This novel procedure presents simple, cheap and stable fabrication strategy for efficient separation of proteins.

HSA immobilized poly(2-hydroxyethylmethacrylate), HSA-P[HEMA], cryogel monolith was examined as an alternative sorbent to be used in hemoperfusion columns for bilirubin removal from serum. The cryogel monolith synthesis was performed by free radical polymerization using MBAA as crosslinker, APS and TEMED as redox pair. Cyanogen bromide (CNBr) was used as a matrix-activating agent for the preparation of immobilized cryogels. Control cryogel monolith, P[HEMA], and HSA-P[HEMA] were characterized by swelling test, SEM images, porosity and surface area measurements, and blood compatibility tests. Activation and immobilization processes were optimized. The removal of bilirubin from plasma samples overloaded with bilirubin was performed using P[HEMA] cryogel monoliths containing different amounts of immobilized HSA in continuous mode. Several factors affecting adsorption capacity of the matrix such as incubation time, HSA concentration, bilirubin concentration in plasma and temperature were analysed. The maximum bilirubin removal from plasma was 25.4 mg/g at 37.5 °C. The desorption agent was 0.1 M NaOH and 1.0 M NaCl containing solution. The reusability was tested for 10 consecutive adsorption–desorption cycles. The adsorption isotherm models and kinetics of process were also studied.

Trietazine was selectively separated from aqueous solution containing the competitor molecule cyanazine, which is similar in size and shape to the template molecule. Structural features of the molecularly imprinted column were figured out by SEM. The influence of the mobile-phase composition, applied electrical field, and pH of the mobile phase on the recognition of trietazine by the imprinted monolithic polymer has been evaluated, and the imprint effect in the trietazine-imprinted monolithic polymer was demonstrated by an imprinting factor. The optimized monolithic column resulted in separation of trietazine from a structurally related competitor molecule, cyanazine. In addition, fast separation was obtained within 6 min by applying higher electrical field, with the electrophoretic mobility of 2.97 × 10(-8) m(2) V(-1) s(-1) at pH 11.0.

A dopamine-imprinted monolithic column was prepared and used in capillary electrochromatography as stationary phase for the first time. Dopamine was selectively separated from aqueous solution containing the competitor molecule norepinephrine, which is similar in size and shape to the template molecule. Morphology of the dopamine-imprinted column was observed by scanning electron microscopy. The influence of the organic solvent content of mobile phase, applied pressure and pH of the mobile phase on the recognition of dopamine by the imprinted monolithic column has been evaluated, and the imprinting effect in the dopamine-imprinted monolithic polymer was verified. Developed dopamine-imprinted monolithic column resulted in excellent separation of dopamine from structurally related competitor molecule, norepinephrine. Separation was achieved in a short period of 10 min, with the electrophoretic mobility of 5.81x10(-5)m(2)V(-1)s(-1) at pH 5.0 and 500mbar pressure.

Myoglobin is a primary iron, and oxygen-binding protein of muscle tissues and levels can be an important diagnostic biomarker for acute myocardial infarction, myocardial necrosis, or other cardiac diseases. The establishment of myoglobin recognition systems is important because of its protein's structural and functional values in physiology, biochemistry, and diagnostic value in some damaged muscle tissue and cardiac diseases. For this purpose, we used molecular imprinting technique for myoglobin recognition from aqueous solutions and human plasma. In the first step, myoglobin-imprinted poly(hydroxyethyl methacrylate) (PHEMA) cryogels (MGb-MIP) were prepared, and optimum myoglobin adsorption conditions were determined. Selectivity experiments have been done with the competitive proteins, and myoglobin adsorption from IgG and albumin-free human plasma was studied. The purity of the desorbed samples was determined with SDS-PAGE. The desorption efficiency and reusability of the MGb-MIP cryogels were tested, and it was shown that without any significant loss in the adsorption capacity, MGb-MIP cryogels can be used a number of times for myoglobin recognition and separation.

A new reactive capillary monolith as an alternative to the commonly employed glycidyl methacrylate-based stationary phases in capillary electrochromatography (CEC) applications was synthesized and post-functionalized with charge-bearing groups. For this purpose, a hydrophilic capillary monolith with reactive 3-chloro-2-hydroxypropyl moiety was first obtained by the copolymerization of a new methacrylate-based monomer, 3-chloro-2-hydroxypropyl methacrylate (HPMA-Cl), with a methacrylic crosslinking agent, ethylene glycol dimethacrylate (EDMA). The presence of 3-chloro-2-hydroxypropyl functionality in poly(HPMA-Cl-co-EDMA) monolith allowed the synthesis of monoliths carrying strongly ionizable anionic, cationic or neutral groups such as sulfonic acid, quaternary ammonium or octadecyl, respectively, via simple and single-stage reactions. In the present study, a reactive poly(HPMA-Cl-co-EDMA) capillary monolith was functionalized with sodium bisulfite (NaHSO3) to have a strongly ionizable sulfonic acid group on the monolith. The resulting monolith providing cathodic electroosmotic flow in CEC was successfully used for the separation of phenol derivatives. The theoretical plate numbers up to 63,000 plates/column were achieved. The results showed that a new promising, reactive support that could be functionalized with different chromatographic ligands for different chromatographic applications was obtained in the study.

The methanol extracts of Astragalus plumosus var. krugianus Chamb. & Matthews afforded sixteen cycloartane glycosides among which krugianoside A, was never reported before. All compounds were evaluated for their cytotoxic activity in human skin fibroblast WS1 cells. For compounds exhibiting no significant effect on WS1 viability, the antioxidant potential was examined. Compounds 1 and 8 prevented elevation of ROS induced by t-BOOH, suggesting the potential activity of these compounds to protect fibroblasts from oxidative stress.

WOS: 000323591100008, PubMed ID: 23896429, Hyaluronic acid (HA) has been used in many applications such as pharmaceutical, clinical and cosmetics, so its separation and purification is very important. In this study, firstly D-glucuronic acid imprinted polymers (MIPs) have been synthesized for the separation of HA which has glucuronic acid part in its structure. MIP particles have characterized by elemental analysis, Fourier Transform Infrared Spectroscopy (FT-IR) and swelling tests. Then, synthesized MIP particles have embedded into polyacrylamide based cryogel. Cryogel has prepared by free radical cryogelation process initiated by N,N,N',N'-Tetramethylethylenediamine (TEMED) and ammonium persulfate CAPS) as redox initiators. This cryogel material was characterized by FT-IR, swelling tests, scanning electron microscopy (SEM) and surface adsorption analyze including pore size analyzer (BET) method. The adsorption of HA has investigated by spectrophotometric method using MIPs embedded into cryogel columns (GAIPEC) and the maximum HA adsorption capacity was found to be 318 mgg(-1). The selectivity of GAIPEC column has estimated using N-acetylglucose amine as interfering agent since this molecule is a part of HA and the results have shown that GAIPEC has been nearly 35 times selective for HA than N-acetylglucose amine. The optimum chromatographic conditions for separation of HA were investigated. pH 7.0 buffer solution for elution and 0.1 M of NaCl solution as desorption agent were used at 0.5 mL min(-1) flow rate. Also, recovery of GAIPEC was investigated and the results have shown that GAIPEC could be used many times without decreasing its adsorption capacity significantly. Here in, combining selectivity of MIP particles and mechanical properties of cryogel, a rigid and stable material was prepared for the separation and purification of HA. To point out this, HA has been isolated from fish eye and fermentation of Streptococcus equi RSKK 679 cell culture. After that, it has characterized and Fast Protein Liquid Chromatography (FPLC) applications have been investigated (C) 2013 Elsevier B.V. All rights reserved.

A new type of polymethacrylate-based monolithic column with chiral stationary phase was prepared for the enantioseparation of aromatic amino acids, namely D,L-phenylalanine, D,L-tyrosine, and D,L-tryptophan by CEC. The monolithic column was prepared by in situ polymerization of butyl methacrylate (BMA), N-methacryloyl-L-histidine methyl ester (MAH), and ethylene dimethacrylate (EDMA) in the presence of porogens. The porogen mixture included DMF and phosphate buffer. MAH was used as a chiral selector. FTIR spectrum of the polymethacrylate-based monolith showed that MAH was incorporated into the polymeric structure via in situ polymerization. Some experimental parameters including pH, concentration of the mobile phase, and MAH concentration with regard to the chiral CEC separation were investigated. Single enantiomers and enantiomer mixtures of the amino acids were separately injected into the monolithic column. It was observed that L-enantiomers of aromatic amino acids migrated before D-enantiomers. The reversal enantiomer migration order for tryptophan was observed upon changing of pH. Using the chiral monolithic column (100 μm id and 375 μm od), the best chiral separation was performed in 35:65% ACN/phosphate buffer (pH 8.0, 10 mM) with an applied voltage of 12 kV in CEC. SEM images showed that the chiral monolithic column has a continuous polymeric skeleton and large through-pore structure.

As alternative hydrophobic adsorbent for DNA adsorption, supermacroporous cryogel disks were synthesized via free radical polymerization. In this study, we have prepared two kinds of cryogel disks: (i) poly(2-hydroxyethyl methacrylate-N-methacryloyl-l-tryptophan) [p(HEMA-MATrp)] cryogel containing specific hydrophobic ligand MATrp; and (ii) monosize p(HEMA-MATrp) particles synthesized via suspension polymerization embedded into p(HEMA) cryogel structure to obtain p(HEMA-MATrp)/p(HEMA) composite cryogel disks. These cryogel disks containing hydrophobic functional group were characterized via swelling studies, Fourier transform infrared spectroscopy, elemental analysis, surface area measurements and scanning electron microscopy. DNA adsorption onto both p(HEMA-MATrp) cryogel and p(HEMA-MATrp)/p(HEMA) composite cryogels was investigated. Maximum adsorption of DNA on p(HEMA-MATrp) cryogel was found to be 15 mg/g polymer. Otherwise, p(HEMA-MATrp)/p(HEMA) composite cryogels significantly increased the DNA adsorption capacity to 38 mg/g polymer. Composite cryogels could be used repeatedly without significant loss on adsorption capacity after 10 repetitive adsorption–desorption cycles. Copyright © 2013 John Wiley & Sons, Ltd.

Poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) [poly(HEMA-GMA)] cryogel was synthesized by cryopolymerization technique at frozen temperature. Iminodiacetic acid (IDA) was then attached covalently to the cryogel as a chelating agent. Then, poly(HEMA-GMA)-IDA cryogel was chelated with Ni(II) ions and this novel metal affinity support was used for adsorption of urease from its aqueous solution. Urease adsorption experiments were carried out in a continuous system by using a peristaltic pump. Maximum urease adsorption onto poly(HEMA-GMA)-IDA-Ni(II) cryogel was found to be 11.30 mg/g cryogel at pH 5.0 acetate buffer and in 25 °C medium temperature. Urease adsorption capacity decreased with increasing ionic strength and increasing chromatographic flow rate. Adsorption kinetics of urease onto poly(HEMA-GMA)-IDA-Ni(II) cryogel was also investigated and it was found that Langmuir adsorption model is applicable for this adsorption study. This novel immobilized metal affinity chromatography support was used 10 times without any decrease at their adsorption capacity. It was also observed that urease enzyme was repeatedly adsorbed and desorbed without significant lost in enzymatic activity.

WOS: 000320895900004, In this study, we applied the epitope imprinting approach to prepare molecularly imprinted monolithic cryogels for immunoglobulin G (IgG) recognition. In this respect, we imprinted F-ab fragments of IgG molecules instead of intact protein molecules via two different non-covalent interactions. In the first approach, we directly coordinated F-ab fragments with N-methacryloyl-L-histidine (MAH), polymerizable derivative of L-histidine, but for the second, we used cupric ions [Cu(II)] as mediator between MAH and F-ab fragments. The monolithic cryogels were characterized by Fourier transform infrared (FTIR) spectroscopy, swelling test, and scanning electron microscopy. Then, the monolithic cryogels were used for F-ab fragment adsorption from aqueous solution while evaluating the factors such as pH and F-ab fragment concentration affecting on adsorption process in continuous set-up. After that, monolithic cryogels were used for IgG adsorption by varying pH, IgG concentration, flowrate, and temperature in appropriate ranges. Maximum IgG adsorption capacities were determined as 32.4 mg/g and 49.0 mg/g for directly coordinated cryogel (MIPDirect) and Cu(II) assisted cryogel (MIPCu(II) (assisted)), respectively. Non-imprinted monolithic cryogels were also prepared for control purposes. In addition to F-ab fragments and IgG molecules, albumin and F-c fragment of IgG molecules were used as competitor biomolecules in order to investigate the selectivity gained by imprinting process. Relative selectivity constants were calculated as 1.47, 2.64 and 3.89 for MIPDirect and 2.90, 8.98, and 11.51 for MIPCu(II) (assisted) for F-ab/IgG, F-ab/F-c, and F-ab/albumin as biomolecule pairs, respectively. The desorption efficiency and reusability of MIPCu(II) assisted cryogel were better than that of MIPDirect. The results reported here showed that the metal ion assistance improved the selectivity features of the imprinted cryogels and allowed to study under milder conditions with enhanced adsorptive properties

WOS: 000404553900191, PubMed: 28608810, Identification of pathogenic microorganisms by traditional methods is slow and cumbersome. Therefore, the focus today is on developing new and quicker analytical methods. In this study, a Surface Plasmon Resonance (SPR) sensor with a microcontact imprinted sensor chip was developed for detecting Salmonella paratyphi. For this purpose, the stamps of the target microorganism were prepared and then, microcontact S. paratyphi-imprinted SPR chips were prepared with the functional monomer N-methacryloyl-L-histidine methyl ester (MAH). Characterization studies of the SPR chips were carried out with ellipsometry and scanning electron microscopy (SEM). The real-time Salmonella paratyphi detection was performed within the range of 2.5 x 10(6)-15 x 10(6) CFU/mL. Selectivity of the prepared sensors was examined by using competing bacterial strains such as Escherichia coli, Staphylococcus aureus and Bacillus subtilis. The imprinting efficiency of the prepared sensor system was determined by evaluating the responses of the SPR chips prepared with both molecularly imprinted polymers (MIPs) and non-imprinted polymers (NIPs). Real sample experiments were performed with apple juice. The recognition of Salmonella paratyphi was achieved using these SPR sensor with a detection limit of 1.4 x 10(6) CFU/mL. In conclusion, SPR sensor has the potential to serve as an excellent candidate for monitoring Salmonella paratyphi in food supplies or contaminated water and clearly makes it possible to develop rapid and appropriate control strategies.

ObjectivesTo analyse the influence of geolocation and ethnicity on the clinical presentation of primary Sjögren's syndrome (SjS) at diagnosis.MethodsThe Big Data Sjögren Project Consortium is an international, multicentre registry designed in 2014. By January 2016, 20 centres from five continents were participating. Multivariable logistic regression analyses were performed.ResultsWe included 7748 women (93%) and 562 men (7%), with a mean age at diagnosis of primary SjS of 53 years. Ethnicity data were available for 7884 patients (95%): 6174 patients (78%) were white, 1066 patients (14%) were Asian, 393 patients (5%) were Hispanic, 104 patients (1%) were black/African-American and 147 patients (2%) were of other ethnicities. SjS was diagnosed a mean of 7 years earlier in black/African-American compared with white patients; the female-to-male ratio was highest in Asian patients (27:1) and lowest in black/African-American patients (7:1); the prevalence of sicca symptoms was lowest in Asian patients; a higher frequency of positive salivary biopsy was found in Hispanic and white patients. A north-south gradient was found with respect to a lower frequency of ocular involvement in northern countries for dry eyes and abnormal ocular tests in Europe (OR 0.46 and 0.44, respectively) and Asia (OR 0.18 and 0.49, respectively) compared with southern countries. Higher frequencies of antinuclear antibodies (ANAs) were reported in northern countries in America (OR=1.48) and Asia (OR=3.80) while, in Europe, northern countries had lowest frequencies of ANAs (OR=0.67) and Ro/La (OR=0.69).ConclusionsThis study provides the first evidence of a strong influence of geolocation and ethnicity on the phenotype of primary SjS at diagnosis.