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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.

Son yıllarda, canlılar ve halk sağlığı için önemli bir tehdit oluşturan fenol ve bileşiklerinin tespit edilmesi önem kazanmaktadır. Bu çalışmada, fenolün tespiti için moleküler baskılama yöntemi temel alınarak yüzey plazmon rezonans (SPR) sensörler tasarlanmıştır. Fenol moleküllerine ait boşluklara sahip polimerik film hazırlanarak UV polimerizasyonu ile SPR sensör yüzeyinden oluşturulmuştur. 0.15 ila 10 nM arasındaki tayin aralığında en düşük saptama sınırı 0.011 nM’dir. Ayrıca, biz katekolün yarışmacı ajan olarak seçicilik deneylerini gerçekleştirdik. Genel olarak, moleküler baskılama yaklaşımı ile hazırlanan fenol baskılanmış SPR sensörünün fenol için oldukça hassas ve seçici olduğu bulunmuştur. Fenol baskılı SPR sensörleri, yüksek seçicilikleri, tekrarlayan kullanımları ve hızlı yanıtları özelliği ile mevcut fenol belirleme yöntemlerine yeni bir yöntem olarak kullanılmak düşünülmektedir.

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.

Increasing of the world population and food demand has led to the development of new agricultural practices to improve food production using more effective pesticides. The controlled release formulations of pesticides are used to delay the release of the biologically active components into their environment for a defined period of time. In these formulations, compounds are incorporated in a polymer matrix by different chemical or physical incorporation techniques. Reducing the toxicity of the compounds and unwanted side effects on non-target organisms are prominent characteristics of these formulations. The development of an ideal controlled release pesticide formulation is just as difficult as developing the ideal pesticide. For these reasons, further research works are needed. In this chapter, the recent applications of sensor systems to detect pesticides in environmental studies with particular attention to the fate of pesticides once introduced in water and soil, and also the advantages of their use are discussed. Current trends in the development of sensor technology for rapid assessment of the environment, as well as challenges for practical implementation and future research directions, are discussed. Findings show that the use of sensors to detect pesticides can improve the quality of the environment and help detect polluted sites.

In this study, copper (II) ion (Cu+2) imprinted poly(hydroxyethyl methacrylate-N-metacryloyl-(L)-cysteine methyl ester [PHEMAC-Cu+2] nanoparticles were synthesized by two-phase mini-emulsion polymerization method and applied to the SPR sensor chip surface for the selective determination of the Cu+2 ions in both aqueous solution, Cu+2-spiked artificial urine and physiological serum samples to investigate the effects of metabolite residues during the analysis. The non-imprinted [PHEMAC] nanoparticles were synthesized by applying the same procedure for the [PHEMAC-Cu+2] nanoparticle synthesis except the addition of Cu+2 ions as a control experiment to evaluate the selectivity of the [PHEMAC-Cu+2] nanoparticles. Roughness differences between [PHEMAC-Cu+2] and [PHEMAC] nanoparticles showed that the imprinting process of Cu+2 ions was performed successfully. [PHEMAC-Cu+2] and [PHEMAC] SPR biosensors prepared by attaching nanoparticles onto the surface of sensor chips, were characterized by atomic force microscope, ellipsometer, contact angle measurements. Langmuir adsorption model was found the most applicable model for this affinity system. Results showed that Cu+2 affinity regions on the surface of [PHEMAC-Cu+2] SPR biosensor were homogeneously distributed and have a monolayer structure. Having the high imprinting efficiency with the imprinting factor of 4.74, the [PHEMAC-Cu+2] SPR biosensor was found to show more selectivity towards the target Cu+2 than the non-imprinted [PHEMAC] SPR biosensor. The selectivity studies of [PHEMAC-Cu+2] SPR biosensors for Cu+2 detection were investigated by using Zn+2 and Ni+2 solutions selected as competitor molecules. The results of intraday and interday precision studies were carried out to ascertain the reproducibility of the proposed method and reported as percent relative standard deviation (%RSD) value.

In this study, we have reported a novel wastewater treatment technique by phenol imprinted bacterial cellulose (BC-MIP) nanofibres with high specificity and adsorption capacity. N-methacryloyl-(L) phenylalanine methyl ester (MAPA) functional monomer was used to create specific binding sites for the template molecule phenol via electrostatic and hydrophobic interactions. BC-MIP nanofibres were synthesized by surface imprinting approach in the presence of different amounts of total monomer (% weight), monomer/template ratio and polymerization time. Then, the nanofibres were characterized by FTIR-ATR, surface area analysis (BET), elemental analysis, scanning electron microscopy (SEM) and contact angle measurements. Adsorption studies were performed with respect to pH, temperature and ionic strength, and the adsorption capacity was calculated by using the spectrophotometer. In order to desorb the adsorbed phenol from BC-MIP nanofibres, 0.1 M NaCl solution was used. Besides, BC-MIP nanofibres were applied to real wastewater samples from Ergene basin in Turkey. The suitable equilibrium isotherm was determined as Langmuir isotherm. To evaluate the selectivity of the BC-MIP nanofibres, similar molecules were utilized as competitor molecules, which were 2-chlorophenol, 4-chlorophenol and 2,4-dichlorophenol. Electrostatic interactions were found to contribute to the generation of specific recognition binding sites. The results have shown that imprinting of phenol was achieved successfully with high adsorption capacity. The phenol removal efficiency was reported up to 97%. BC-MIP nanofibres were used 10 times with a negligible decrease in adsorption capacity. [GRAPHICS] . WOS:000471539100001 2-s2.0-85065257535 PubMed: 30919740

The ability to detect catecholamines (CAs) and their metabolites is vital to understand the mechanism behind the neuronal diseases. Neurochemistry aims to provide an improved pharmacological, molecular and physiological understanding of complex brain chemistries by analytical techniques. Capillary electrophoresis (CE) is one such analytical technique that enables the study of various chemical species ranging from amino acids and peptides to natural products and drugs. CE can easily adapt the changes in research focus and in recent years remains an applicable technique for investigating neuroscience and single cell neurobiology. The prepared phenylalanine-based hydrophobic monolithic column, Polymethacryloyl-L-phenylalanine [PMAPA], was used as a stationary phase in capillary electrochromatography to separate CAs that are similar in size and shape to each other including dopamine (DA) and norepinephrine (NE) via hydrophobic interactions. Separation carried out in a short period of 17 min was performed with the electrophoretic mobility of 5.54 × 10−6 m2 V−1 s−1 and 7.60 × 10−6 m2 V−1 s−1 for DA and NE, respectively, at pH 7.0, 65% acetonitrile ratio with 100 mbar applied pressure by the developed hydrophobic monolithic column without needing any extra process such as imprinting or spacer arms to immobilize ligands used in separation.

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.

One-step activation, purification, and stabilization of lipase enzyme were performed by using composite hydrophobic support at low ionic strength with increased surface area during embedding process. A novel hydrophobic poly(hydroxyethylmethacrylate) [PHEMA] based, poly(glycidyl methacrylate-N-methacryloly-(L)-tryptophan) [PGMATrp] bead embedded composite cryogel membrane having specific surface area of 195 m(2)/g was used as hydrophobic matrix for adsorption of commercial Candida Rugosa lipase in a continuous system. PGMATrp embedded PHEMA cryogel membrane with 60-100 mu m pore size was obtained by dispersion polymerization of GMA and MATrp to form PGMATrp beads followed by embedding of PGMATrp to HEMA via APS and TEMED redox pair. The introduction of hydrophobic MATrp monomer into bead structure aiming to increase interaction between lipase and composite membrane was estimated using nitrogen stoichiometry of elemental analysis and found to be 239 mu mol/g of polymer. Hydophobicity increment due to embedding process was confirmed by measuring contact angle, it was found 42 degrees and 48.4 degrees for the PHEMA and PHEMA/PGMATrp composite cryogel respectively. Some parameters i.e. pH, flow-rate, protein concentration, temperature, salt type and ionic intensity were evaluated on the adsorption capacity in a continuous system. Fast protein liquid chromatography (FPLC) studies were performed for specific adsorption of lipase onto the PHEMA/PGMATrp embedded composite cryogel membrane. (C) 2015 Elsevier B.V. All rights reserved.

The use of molecular imprinted polymers (MIPs), which provides a means for preparing stationary phases with predetermined selectivity for a target molecule in capillary electrochromatography (CEC), is attractive because it combines selectivity, higher separation efficiency and shorter analysis time. A bisphenol A (BPA)-imprinted monolithic capillary BPA/PMAPA column was synthesized for the selective separation of bisphenol A (BPA) from aqueous solutions containing the competitor molecule phenol (PH), which is similar in size and shape to the template molecule. BPA-imprinted monolithic column was prepared in the presence of the template molecule, BPA, which results in the formation of recognition cavities complementary to the template molecule, after the removal of template molecule. An amino acid based monomer, N-methacryloyl-L-phenyl alanine (MAPA), was used as the functional monomer. The new stationary phase contains both charged and hydrophobic groups originating from MAPA monomer, which behaves as both an electroosmotic flow (EOF) supplier and a hydrophobic matrix. The MAPA containing BPA imprinted column behaves as a mixed mode stationary phase, as ion exchanger and hydrophobic matrix depending on the pH of the medium. Scanning electron microscopy was used to identify the structural features of the molecular imprinted column. MIP column performance was evaluated by the electrochromatographic separation of alkylbenzenes. The novelty of this work originated from dual separation mechanism shown by MAPA, which has the ability to form both hydrophobic and electrostatic interactions by the charged and hydrophobic groups of phenylalanine amino acid. This new column with mixed-mode characteristics was then used successfully as the stationary phase in CEC for the selective separation of BPA in MIP systems.

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.

Pesticides have been utilized in agriculture for decades. However, their widespread use has increased multiple concerns due to their known and suspected toxicities on long-term human health risks in scientific and industrial communities. Thus, detecting pesticides will have a great impact on their management, as well as improve their toxicity effects over humans. Here, we fabricate molecularly imprinted nanofilms and integrate them with surface plasmon resonance (SPR) sensors for sensitive, selective, fast and real-time detection of multiple pesticides, including cyanazine (SNZ), simazine (SMZ) and atrazine (ATZ). The molecularly imprinted nanofilms onto the SPR gold surfaces are prepared via UV polymerization reactions, which consist of N-methacryloyl-L-phenylalanine methyl ester (MAPA) as a functional monomer, 1-vinylimidazole (VIM) as a co-monomer, and ethylene glycol dimethacrylate (EGDMA) as a cross-linker. The real-time measurements on SPR sensor provide a detection range from 0.10 to 6.64 nM, as well as denote a limit of detection (LOD) values of 0.095, 0.031 and 0.091 nM for SNZ, SMZ and ATZ, respectively. Furthermore, we perform selectivity test, where SNZ, SMZ and ATZ are examined as competitor agents. Overall, the pesticide imprinted SPR sensors have been found to be highly selective and sensitive. These SPR sensors also hold great potential to be used an alternative method for the existing pesticide monitoring approaches due to their reusability, fast response, and easy-to-use properties, as well as can be tailored to detect and real-time monitor of other pesticides. (C) 2016 Elsevier B.V. All rights reserved.

A molecularly imprinted composite cryogel (MICC) was prepared for depletion of hemoglobin from human blood prior to use in proteome applications. Poly(hydroxyethyl methacrylate) based MICC was prepared with high gel fraction yields up to 90%, and characterized by Fourier transform infrared spectrophotometer, scanning electron microscopy, swelling studies, flow dynamics and surface area measurements. MICC exhibited a high binding capacity and selectivity for hemoglobin in the presence of immunoglobulin G, albumin and myoglobin. MICC column was successfully applied in fast protein liquid chromatography system for selective depletion of hemoglobin for human blood. The depletion ratio was highly increased by embedding microspheres into the cryogel (93.2%). Finally, MICC can be reused many times with no apparent decrease in hemoglobin adsorption capacity. Copyright © 2014 John Wiley & Sons, Ltd.

Herein, we have focused on the preparation of triazinic pesticide imprinted SPR nanosensors for detection of herbicides. Triazinic pesticides are weedkillers that are related with possible carcinogenic effects, birth defects, and menstrual problems when uptake by humans. Although there are restrictions and bans on their use in some countries they are still one of the most widely used pesticides in the world. The development of rapid, sensitive, and inexpensive diagnosis tools for environmental and biological monitoring is currently a research area of great interest. Surface plasmon resonance (SPR) nanosensors have been used widely for the detection of triazinic pesticides because of their simplicity, lack of requirement for labeling and ease of miniaturization, low cost, high specificity and sensitivity, and real-time measurement. Molecularly imprinted polymers that have molecular recognition talent, are easy to prepare, less expensive, stable, and can be manufactured with good reproducibility, are used for the creation of biorecognitive surfaces on the SPR nanosensors. Herein, we have focused on the production of triazinic pesticide-imprinted SPR nanosensors.

WOS: 000407478900004, PubMed: 28322473, Diagnostic biomarkers such as proteins and enzymes are generally hard to detect because of the low abundance in biological fluids. To solve this problem, the advantages of surface plasmon resonance (SPR) and nanomaterial technologies have been combined. The SPR sensors are easy to prepare, no requirement of labelling and can be detected in real time. In addition, they have high specificity and sensitivity with low cost. The nanomaterials have also crucial functions such as efficiency improvement, selectivity, and sensitivity of the detection systems. In this report, an SPR-based sensor is developed to detect lysozyme with hydrophobic poly (N-methacryloyl-(L)-phenylalanine) (PMAPA) nanoparticles. The SPR sensor was first characterized by attenuated total reflection-Fourier transform infrared, atomic force microscope, and water contact angle measurements and performed with aqueous lysozyme solutions. Various concentrations of lysozyme solution were used to calculate kinetic and affinity coefficients. The equilibrium and adsorption isotherm models of interactions between lysozyme solutions and SPR sensor were determined and the maximum reflection, association, and dissociation constants were calculated by Langmuir model as 4.87, 0.019nM(-1), and 54nM, respectively. The selectivity studies of SPR sensor were investigated with competitive agents, hemoglobin, and myoglobin. Also, the SPR sensor was used four times in adsorption/desorption/recovery cycles and results showed that, the combination of optical SPR sensor with hydrophobic ionizable PMAPA nanoparticles in one mode enabled the detection of lysozyme molecule with high accuracy, good sensivity, real-time, label-free, and a low-detection limit of 0.66nM from lysozyme solutions. Lysozyme detection in a real sample was performed by using chicken egg white to evaluate interfering molecules present in the medium.

The immobilized metal-affinity chromatography (IMAC) has gained significant interest as a widespread separation and purification tool for therapeutic proteins, nucleic acids and other biological molecules. The enormous potential of IMAC for proteins with natural surface exposed-histidine residues and for recombinant proteins with histidine clusters. Cryogels as monolithic materials have recently been proposed as promising chromatographic adsorbents for the separation of biomolecules in downstream processing. In the present study, IMAC cryogels have been synthesized and utilized for the adsorption and separation of immunoglobulin G (IgG) from IgG solution and whole human plasma. For this purpose, Cu(II)-ions were coupled to poly(hydroxyethyl methacrylate) PHEMA using poly(ethylene imine) (PEI) as the chelating ligand. In this study the cryogels formation optimized by the varied proportion of PEI from 1% to 15% along with different amounts of Cu (II) as chelating metal. The prepared cryogels were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The [PHEMA/PEI]-Cu(II) cryogels were assayed for their capability to bind the human IgG from aqueous solutions. The IMAC cryogels were found to have high affinity toward human IgG. The adsorption of human IgG was investigated onto the PHEMA/PEI cryogels with (10% PEI) and the concentration of Cu (II) varied as 10, 50,100 and 150 mg/L. The separation of human IgG was achieved in one purification step at pH 7.4. The maximum adsorption capacity was observed at the [PHEMA/PEI]-Cu(II) (10% PEI) with 72.28 mg/g of human IgG. The purification efficiency and human IgG purity were investigated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). (C) 2016 Elsevier B.V. All rights reserved.

Glycoamylase (AMG) is an ?-amylase enzyme which catalyzes the breakdown of large a(1,4)-linked malto-oligosaccharides to glucose. It is an extracellular enzyme and is excreted to the culture medium. In this study, AMG was immobilized on a variety of metal affinity membranes, which were prepared by chelating Cu2+ ions onto poly(hydroxyethyl methacrylate) (PHEMA) using N-methacryloyl-(L)-histidine methyl ester (MAH), N-methacryloyl-(L)-cysteine methyl ester (MAC), and N-methacryloyl-(L)-phenylalanine methyl ester (MAPA) as metal-chelating comonomers for reversible immobilization of AMG. The PHEMAH, PHEMAC, PHEMAPA membranes were synthesized by UV-initiated photo-polymerization and Cu2+ ions were chelated on the membrane surfaces. Cu2+-chelated membranes were characterized by swelling tests, SEM, contact angle measurements, elemental analysis, and FTIR. AMG immobilization on the Cu2+-chelated membranes was performed by using aqueous solutions of different amounts of AMG at different pH values and Cu2+ loadings. Durability tests concerning desorption of AMG and reusability of the Cu2+-chelated membranes yielded acceptable results. It was computationally determined that AMG possesses four likely Cu2+/Zn2+ binding sites, away from the catalytic site, to which the metal-chelated membranes can be efficiently used. (C) 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

WOS: 000276436300007, A new generation of molecularly imprinted macroporous hydrogels, so-called cryogels, were prepared for depletion of hemoglobin from human blood. Hb-PHEMAH cryogels were prepared with gel fraction yields up to 90%, and its morphology and porosity were characterized by FTIR, SEM, swelling studies, flow dynamics and surface area measurements. Selective binding experiments were performed in the presence of competitive PHEMAH cryogel exhibited a high binding capacity and selectivity for Hb in the presence of Myb and BSA. The selectivity of Hb-PHEMAH cryogel for Hb was confirmed by HPLC. Hb depletion from blood hemolysate was also studied using SDS-PAGE. Hb-PHE-MAH cryogel can be reused many times with no apparent decrease in Hb adsorption capacity.

WOS: 000257442900022, Supermacroporous poly(2-hydroxyethyl methacrylate) [poly(HEMA)]-based monolithic cryogel column was prepared by radical cryocopolymerization of HEMA with N-methacryloyl-L-histidine methyl ester (MAH) as functional comonomer and N,N'-methylene-bisacrylamide (MBAAm) as crosslinker directly in a plastic syringe for affinity purification of lysozyme from chicken egg white. The monolithic cryogel containing a continuous polymeric matrix having interconnected pores of 10-50 mu m size was loaded with Zn2+ ions to form the metal chelate with poly(HEMA-MAH) cryogel. Poly(HEMA-MAH) cryogel was characterized by swelling studies, FTIR, scanning electron microscopy, and elemental analysis. The equilibrium swelling degree of the poly(HEMA-MAH) monolithic cryogel was 5.62 g H2O/g cryogel. Poly(HEMA-MAH) cryogel containing 45.8 mu mol MAH/g was used in the adsorption/desorption of lysozyme from aqueous solutions. The nonspecific adsorption of lysozyme was very low (7.5 mg/g). The maximum amount of lysozyme adsorption from aqueous solution in phosphate buffer was 209 mg/g at pH 7.0. It was observed that lysozyme could be repeatedly adsorbed and desorbed with the poly (HEMA-MAH) cyogel without significant loss of adsorption capacity. (C) 2008 Wiley Periodicals, Inc.

We have synthesized PVA/PEI monolithic cryogel columns chelated with Cu2+ ions as a model adsorbent, which is capable of binding haemoglobin (Hb) from human blood. The goal of this study is to perform the depletion of Hb via a single and easy process to be useful in proteomic studies. PVA/PEI-Cu2+ cryogel columns were subjected to adsorption studies of Hb from both aqueous solution and human plasma to evaluate the extent of interaction between cryogel columns and Hb. The effects of experimental parameters, such as pH, Hb equilibrium concentration, adsorption time, temperature, and ionic strength, on Hb adsorption capacity were investigated.

WOS: 000376136500005, PubMed: 26772759, In this study, we aimed to prepare the metal chelate affinity cryogels for the hemoglobin (Hb) depletion. Poly(2-hydroxyethyl methacrylate) (PHEMA) cryogels were selected as base matrix because of their blood compatibility, osmotic, chemical, and mechanical stability. Cryogels are also useful when working with the viscous samples such as blood, because of their interconnected macroporous structure. Iminodiacetic acid (IDA), the chelating agent, was covalently coupled with PHEMA cryogels after activation with the epichlorohydrin and then the Ni(II) ions were chelated to the IDA-bound cryogels. The depletion of the Hb from hemolysate was shown by SDS-PAGE.

Surface plasmon resonance (SPR) sensors have been used for detection of various biomolecules because of their simplicity, high specificity and sensitivity, real-time detection, low cost, and no requirement of labeling. Recently, molecularly imprinted polymers that are easy to prepare, less expensive, stable, have talent for molecular recognition and also are used for creation selective binding sites for target molecule on the SPR sensors. Here, we show that preparation of cyclic citrullinated peptide antibody (anti-CCP) imprinted SPR sensor to detect CCP antibodies. For this purpose, anti-CCP/AAm pre-complex was synthesized by interacting acrylamide (AAm) monomer with anti-CCP. Then, anti-CCP imprinted (anti-CCP/PAAm) SPR sensor was obtained by reacting with anti-CCP/AAm pre-complex in the presence of the crosslinker, and initiator/activator pair. Besides this, non-imprinted (PAAm) SPR sensor was also prepared without using anti-CCP template. The SPR sensors were characterized and then adsorption-desorption studies were performed with pH 7.0 phosphate buffer (10mM) and acetic acid (10%) with Tween 20 (1%) in pH 7.0 phosphate buffer. Selectivitiy of sensors was investigated by using immunoglobulin M (IgM) and bovine serum albumin (BSA). To determine the adsorption model of interactions between anti-CCP solutions and anti-CCP/PAAm SPR sensor, different adsorption models were performed. The calculated maximum reflection, detection limit, association and dissociation constants were 1.079RU/mL, 0.177RU/mL, 0.589RU/mL and 1.697mL/RU, respectively. Repeatability experiments of anti-CCP/PAAm SPR sensor was performed four times with adsorption-desorption-regeneration cycles without any performance losing. Results showed that anti-CCP/PAAm SPR sensor had high selectivity and sensitivity for detection of CCP antibodies.

A molecularly imprinted composite cryogel (MICC) was prepared for depletion of hemoglobin from human blood prior to use in proteome applications. Poly(hydroxyethyl methacrylate) based MICC was prepared with high gel fraction yields up to 90%, and characterized by Fourier transform infrared spectrophotometer, scanning electron microscopy, swelling studies, flow dynamics and surface area measurements. MICC exhibited a high binding capacity and selectivity for hemoglobin in the presence of immunoglobulin G, albumin and myoglobin. MICC column was successfully applied in fast protein liquid chromatography system for selective depletion of hemoglobin for human blood. The depletion ratio was highly increased by embedding microspheres into the cryogel (93.2%). Finally, MICC can be reused many times with no apparent decrease in hemoglobin adsorption capacity.

Immobilized metal ion affinity chromatography (IMAC) is a useful method for adsorption of proteins that have an affinity for transition metal ions. In this study, poly( hydroxyethyl methacrylate-methacryloyl-L-tryptophan) (PHEMATrp) nanoparticles were prepared by surfactant free emulsion polymerization. Then, Cu(II) ions were chelated on the PHEMATrp nanoparticles to be used in lysozyme adsorption studies in batch system. The maximum lysozyme adsorption capacity of the PHEMATrp nanoparticles was found to be 326.9 mg/g polymer at pH 7.0. The nonspecific lysozyme adsorption onto the PHEMA nanoparticles was negligible. In terms of protein desorption, it was observed that adsorbed lysozyme was readily desorbed in medium containing 1.0 M NaCl. The results showed that the metal-chelated PHEMATrp nanoparticles can be considered as a good adsorbent for lysozyme purification. (C) 2012 Elsevier B.V. All rights reserved.

WOS: 000275342200010, DNA adsorption properties of Zn(2+)-chelated supermacroporous poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine methyl ester) [poly(HEMA-MAH)] monolithic cryogel column were investigated for the application of DNA-affinity adsorbents. The monolithic cryogel was loaded with Zn(2+) ions to form the metal-chelated affinity sorbent. Poly(HEMA-MAH) cryogel was characterized by swelling tests, FTIR, scanning electron microscopy (SEM), and elemental analysis. SEM analysis indicates that the cryogel have a heteroporous structure with interconnected pores of 10-50 mu m size, which ascribed to the porogens effect of frozen water crystals. Poly(HEMA-MAH) cryogel containing 45.8 mu mol MAH was used in the adsorption/desorption of DNA from aqueous solutions. The maximum amount of DNA adsorption was 32.93 mg/g in Tris buffer at pH 7.0. It was observed that DNA could be repeatedly adsorbed and desorbed with the poly(HEMA-MAH) cryogel without significant loss of adsorption capacity. As a result, these higher amounts of DNA adsorbed poly (HEMA-MAH) cryogels are expected to be good candidate for achieving higher removal of anti-DNA antibodies from systemic lupus erythematosus (SLE) patients plasma. (C) 2009 Wiley Periodicals, Inc. J Appl Polym Sci 116: 1306-1312, 2010