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In this study, quartz crystal microbalance sensors based on molecular imprinting technology were fabricated for real-time detection of insulin in aqueous solution and artificial plasma. This study describes the preparation of insulin imprinted poly(hydroxyethyl methacrylate)-N-methacryloyl-( l )-histidine methyl ester based quartz crystal microbalance sensor for insulin determination. Poly(hydroxyethyl methacrylate)-N-methacryloyl-( l )-histidine methyl ester based film on chip surface was synthesized by ultra violet (UV) polymerization for the detection of insulin at low concentrations. At the first step, N-methacryloyl-( l )-histidine methyl ester complex was formed with insulin and then, the insulin imprinted film has been prepared. The characterization of the polymeric film has been conducted with ellipsometry, contact angle, Fourier transform infrared-attenuated total reflectance and atomic force microscopy measurements. Langmuir, Freundlich and Langmuir-Freundlich adsorption isotherm models were applied for this system. The best fitted model to explain the interactions between molecular imprinted chip and insulin molecules was the Langmuir adsorption isotherm (R2: 0.999). The repeatability of insulin imprinted chip was investigated by using of equilibration-binding-regeneration cycles for four times. The detection limit was found as 0.00158 ng/mL. According to the results, the QCM sensor has showed low-detection limit, high selectivity and sensitivity for insulin assay.

In this study, cryogel-based molecularly imprinted composite cartridges were designed for the rapid, efficient, and selective preconcentration of benzo[ a ]pyrene (BaP) from water samples. First, a BaP-imprinted poly(2-hydroxyethyl methacrylate-N-methacryloyl-(L)-phenylalanine) composite cartridge was synthesized under semi-frozen conditions and characterized by scanning electron microscopy, elemental analysis, Fourier transform infrared spectroscopy, and swelling tests. After the optimization of preconcentration parameters, i.e., pH and initial BaP concentration, the selectivity and preconcentration efficiency, and reusability of these cartridges were also evaluated. In selectivity experiments, BaP imprinted composite cartridge exhibited binding capacities 3.09, 9.52, 8.87, and 8.77-fold higher than that of the non-imprinted composite cartridge in the presence of competitors, such as benzo[ b ]fluoranthene (BbF), benzo[ k ]fluoranthene (BkF), indeno[1,2,3- cd ]pyrene (IcdP), and 1-naphthol, respectively. The method detection limit (MDL), relative standard deviation (RSD) and preconcentration efficiency (PE) of the synthesized composite cartridge were calculated as 24.86 μg/L, 1.60%, and 349.6%, respectively.

The aim of this study is to prepare megaporous dye-affinity cryogel discs for the purification of plasmid DNA (pDNA) from bacterial lysate. Poly(hydroxyethyl methacrylate) [PHEMA] cryogel discs were produced by free radical polymerization initiated by N,N,N',N'-tetramethylene diamine (TEMED) and ammonium persulfate (APS) redox pair in an ice bath. Cibacron Blue F3GA was used as an affinity ligand (loading amount: 68.9μmol/g polymer). The amount of pDNA adsorbed onto the PHEMA-Cibacron Blue F3GA cryogel discs first increased and then reached a plateau value (i.e., 32.5mg/g cryogel) at 3.0mg/mL pDNA concentration. Compared with the PHEMA cryogel (0.11mg/g cryogel), the pDNA adsorption capacity of the PHEMA-Cibacron Blue F3GA cryogel (32.4mg/g polymer) was improved significantly due to the Cibacron Blue 3GA immobilization onto the polymeric matrix. pDNA adsorption amount decreased from 11.7mg/g to 1.1mg/g with the increasing of NaCl concentration. The maximum pDNA adsorption was achieved at 4°C. The overall recovery of pDNA was calculated as 90%. The PHEMA-Cibacron Blue F3GA cryogel discs could be used five times without decreasing the pDNA adsorption capacity significantly. The results show that the PHEMA-Cibacron Blue F3GA cryogel discs promise high selectivity for pDNA.

Immunoaffinity chromatography (IAC) is a kind of bioaffinity chromatography which used antibodies or antibody-related molecules as the stationary phase. IAC is used by many applications for analytical, clinical and diagnostic purposes, particularly preferring in analytical purposes on one-step separation and purification of target compounds. Moreover, immunoaffinity chromatography is used in antibody enrichment and separation of cells. IAC columns are usually applied in the antibody experiments due to powerful and selective binding of antibodies and/or their target antigens. Antigen or antibody molecules could be immobilized to the solid support. Therefore, target antibody or cell is purified. Specific bioligands can be immobilized directly on glycidyl based polymeric material with simple acid–base catalyst. In this study, polyglycidyl methacrylate based therefore cryogels were prepared and anti-insulin antibodies were immobilized on porous surface of cryogels. Swelling test, Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, and scanning electron microscopy (SEM) were conducted to characterize cryogels developed. To optimize separation conditions, effects of pH, initial insulin concentration, flow rate, salt concentration, contact time and temperature on insulin adsorption capacity were examined. The results indicated that the immunoaffinity cryogel developed here could be classified as good alternative with prominent properties such as high reusability and cost-friendly adsorbent and would be one of the primary reports for immunoaffinity purification of insulin molecules in not only lab-scale but also for industrial purposes.

Naturally produced by the human immune system, immunoglobulin nowadays is widely used for in vivo and in vitro purposes. The increased needs for pure immunoglobulin have prompted researchers to find new immunoglobulin chromatographic separation processes. Cryogels as chromatographic adsorbents, congregate several mechanical features including good compatibility, large pore structure, flexibility, short diffusion pathway and stability. These different characteristics make them a good alternative to conventional chromatographic methods and allowing their potential use in separation technology. In the present study, two sets of poly(2-hydroxyethyl methacrylate) (PHEMA) based beads were prepared and functionalized with Reactive Red 120 (RR) and Reactive Green HE 4BD (RG) dyes, and then embedded into supermacroporous cryogels. The morphology, physical and chemical features of the prepared bead embedded composite cryogel discs (CCDs) were performed by scanning electron microscopy (SEM), swelling test, elemental analysis and Fourier transform infrared spectroscopy (FTIR). The results showed that the embedded composite cryogel discs have a specific surface area of 192.0 m2/g with maximum adsorption capacity of HIgG 239.8 mg/g for the RR functionalized CCD and 170 mg/g for RG functionalized CCD columns, both at pH 6.2.

The aim of this study is to prepare silanized polymeric nanoparticles for DNA isolation. Polymeric p(HEMA)-IMEO-PBA nanoparticles around 85.7 nm diameter, was obtained by surfactant free emulsion polymerization for DNA isolation. Synthesized nanoparticles for characterization studies were realized scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and Zeta-size. Surface area, average particle size and size distribution were also performed. The surface area of synthesized silanized polymeric nanoparticles was 2460 m 2 /g. Synthesized polymeric nanoparticles were silanized with 3-(2-imidazoline-1-yl)propyl (triethoxysilane) (IMEO). After that, phenylboronic acid (PBA) which is DNA specific ligand were covalently binded to silanized polymeric nanoparticles. The amount of DNA adsorbed onto the p(HEMA)-IMEO-PBA nanoparticles first increased and then reached a saturation value at around 14.0 mg/mL of DNA concentration. The maximum adsorption was 672.41 mg/g silanized polymeric nanoparticles in the optimum adsorption medium. The maximum DNA adsorption was achieved at 4 °C. The overall recovery of DNA was calculated as 95%. In repetitive adsorption–desorption circles, it is observed not being important decrease in DNA adsorption capacities. The results were shown that silanized polymeric nanoparticles can be a good alternative for DNA isolation.

Supermacroporous cryogel with an average pore size of 10–200 μm in diameter was prepared by cryopolymerization of N-methacryloyl-( l )-glutamic acid (MAGA) and 2-hydroxyethyl methacrylate (HEMA). The poly(HEMA–MAGA) cryogel was characterized by surface area measurements, FTIR, swelling studies, elemental analysis and SEM. The poly(HEMA–MAGA) cryogel had a specific surface area of 23.2 m 2 /g. The equilibrium swelling ratio of the cryogel is 9.68 g H 2 O/g for poly(HEMA–MAGA) and 8.56 g H 2 O/g cryogel for PHEMA. The poly(HEMA–MAGA) cryogel disks with a pore volume of 71.6% containing 878 μmol MAGA/g were used in the removal of UO 2 2+ from aqueous solutions. Adsorption equilibrium of UO 2 2+ was obtained in about 30 min. The adsorption of UO 2 2+ ions onto the PHEMA cryogel disks was negligible (0.78 mg/g). The MAGA incorporation significantly increased the UO 2 2+ adsorption capacity (92.5 mg/g). The adsorption process is found to be a function of pH of the UO 2 2+ solution, with the optimum value being pH 6.0. Adsorption capacity of MAGA contained PHEMA based cryogel disks increased significantly with pH and then reached the maximum at pH 6.0. Consecutive adsorption and elution cycles showed the feasibility of repeated use for poly(HEMA–MAGA) cryogel disks.

The aim of this study is to prepare poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-histidine) [PHEMAH] magnetic nanoparticles for plasmid DNA (pDNA) purification from Escherichia coli (E. coli) cell lysate. Magnetic nanoparticles were produced by surfactant free emulsion polymerization. mPHEMAH nanoparticles were characterized by elemental analysis, Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), vibrating sample magnetometer (VSM), electron spin resonance (ESR), thermogravimetric analyses (TGA) and transmission electron microscopy (TEM). Surface area, average particle size and size distribution were also performed. Specific surface area of the mPHEMAH nanoparticles was found to be 1180 m2/g. Elemental analysis of MAH for nitrogen was estimated as 0.18 mmol/g polymer. The amount of pDNA adsorbed onto the mPHEMAH nanoparticles first increased and then reached a saturation value at around 1.0 mg/mL of pDNA concentration. Compared with the mPHEMA nanoparticles (50 μg/g polymer), the pDNA adsorption capacity of the mPHEMAH nanoparticles (154 mg/g polymer) was improved significantly due to the MAH incorporation into the polymeric matrix. The maximum pDNA adsorption was achieved at 25 °C. The overall recovery of pDNA was calculated as 92%. The mPHEMAH nanoparticles could be used six times without decreasing the pDNA adsorption capacity significantly. The results indicate that the PHEMAH nanoparticles promise high selectivity for pDNA.

WOS: 000303299300046, Hydrophobic interaction chromatography (HIC) is increasingly used for protein purification, separation and other biochemical applications. The aim of this study was to prepare hydrophobic microbeads and to investigate their recombinant human interferon-alpha (rHuIFN-alpha) adsorption capability. For this purpose, we had synthesized functional monomer, N-methacryloyl-L-phenylalanine (MAPA), to provide a hydrophobic functionality to the adsorbent. The poly(2-hydroxyethyl methacrylate-N-methacryloyl-L-phenylalanine) [poly(HEMA-MAPA)] microbeads were prepared by suspension copolymerization. microbeads were characterized using FTIR, swelling behavior, and SEM micrographs. Equilibrium swelling ratio of poly(HEMA-MAPA) and poly(HEMA) microbeads were 53.3% and 69.3%, respectively. The specific surface area and average pore diameters determined by BET apparatus were 17.4 m(2)/g and 47.3 angstrom for poly(HEMA) microbeads and 18.7 m(2)/g and 49.8 angstrom for poly(HEMA-MAPA) microbeads. Adsorption experiments were performed under different conditions. Maximum rHuIFN-alpha adsorption capacity was found to be 137.6 +/- 6.7 mg/g by using poly(HEMA-MAPA) microbeads with a size range of 150-250 mu m and containing 327 mu mol MAPA/g microbeads. Compared with poly(HEMA-MAPA) microbeads, nonspecific rHuIFN-alpha adsorption onto plain poly(HEMA) microbeads was very low, about 4.2 +/- 2.3 mg/g. To determine the effects of adsorption conditions on possible conformational changes of rHuIFN-alpha structure, fluorescence spectrophotometry was employed. Repeated adsorption-elution processes showed that these microbeads are suitable for repeatable rHuIFN-alpha adsorption

Supermacroporous poly(2-hydroxyethyl methacrylate) (PHEMA) cryogel carrying DNA was used in the removal of anti-dsDNA antibodies from systemic lupus erythematosus (SLE) patient plasma. The PHEMA cryogel was prepared by bulk polymerization which proceeds in an aqueous solution of monomer frozen inside a plastic syringe. After thawing, the PHEMA cryogel contains a continuous matrix having interconnected macropores of 10–200 μm size. Pore volume in the PHEMA cryogel was 67.5%. Ester groups in the PHEMA structure were converted to imine groups by reacting with poly(ethyleneimine) (PEI) in the presence of NaHCO 3 . Amino (− NH 2 ) content of PEI-modified PHEMA cryogel was determined as 82 mg PEI/g. Then, DNA was attached onto the PHEMA cryogel via amino groups (53.4 mg DNA/g cryogel). Anti-dsDNA-antibody concentration declined significantly from 780 IU/ml to 80 IU/ml with the time. The maximum anti-dsDNA-antibody adsorption amount was 70 × 10 3 IU/g. Anti-dsDNA-antibodies could be repeatedly adsorbed and eluted without noticeable loss in the anti-dsDNA-antibody adsorption amount.

Low density lipoprotein (LDL) cholesterol is a major ingredient of the plaque that collects in the coronary arteries and causes coronary heart diseases. Among the methods used for the extracorporeal elimination of LDL from intravasal volume, immunoaffinity technique using anti-LDL antibody as a ligand offers superior selectivity and specificity. Proper orientation of the immobilized antibody is the main issue in immunoaffinity techniques. In this study, anti-human β-lipoprotein antibody (anti-LDL antibody) molecules were immobilized and oriented through protein A onto poly(2-hydroxyethyl methacrylate) (PHEMA) cryogel in order to remove LDL from hypercholesterolemic human plasma. PHEMA cryogel was prepared by free radical polymerization initiated with N,N,N′,N′-tetramethylene diamine (TEMED). PHEMA cryogel with a swelling degree of 8.89 g H 2 O/g and 67% macro-porosity was characterized by swelling studies, scanning electron microscope (SEM) and blood compatibility tests. All the clotting times were increased when compared with control plasma. The maximum immobilized anti-LDL antibody amount was 63.2 mg/g in the case of random antibody immobilization and 19.6 mg/g in the case of oriented antibody immobilization (protein A loading was 57.0 mg/g). Random and oriented anti-LDL antibody immobilized PHEMA cryogels adsorbed 111 and 129 mg LDL/g cryogel from hypercholesterolemic human plasma, respectively. Up to 80% of the adsorbed LDL was desorbed. The adsorption–desorption cycle was repeated 6 times using the same cryogel. There was no significant loss of LDL adsorption capacity.

Novel nanospheres with an average size of 350 nm utilizing N-methacryloyl-( l )-tryptophane methyl ester (MATrp) as a hydrophobic monomer were prepared by surfactant free emulsion polymerization of 2-hydroxyethyl methacrylate (HEMA), (MATrp) conducted in an aqueous dispersion medium. MATrp was synthesized using methacryloyl chloride and ( l )-tryptophane methyl ester. Specific surface area of the non-porous nanospheres was found to be 1902.3 m2/g. poly(HEMA–MATrp) nanospheres were characterized by Fourier Transform Infrared Spectroscopy (FTIR) and scanning electron microscopy (SEM). Average particle size and size distribution measurements were also performed. Elemental analysis of MATrp for nitrogen was estimated at 1.36 mmol/g nanospheres. Then, poly(HEMA–MATrp) nanospheres were used in the adsorption of porcine pancreas lipase in a batch system. Using an optimized adsorption protocol, a very high loading of 558 mg enzyme/g nanospheres was obtained. The adsorption phenomena appeared to follow a typical Langmuir isotherm. The Km value for immobilized lipase (16.26 mM) was higher than that of free enzyme (10.34 mM). It was observed that enzyme could be repeatedly adsorbed and desorbed without significant loss in adsorption amount or enzyme activity. These findings show considerable promise for this material as an adsorption matrix in industrial processes.

Depletion of high abundant proteins like albumin and immunoglobulin G (IgG) can be beneficial in the analysis of serum proteins. For this purpose, Cibacron Blue F3GA and iminodiacetic acid (IDA)-Cu 2+ containing poly(glycidyl methacrylate) (PGMA) beads (1.6 µm in diameter) were embedded into the poly(hydroxyethyl methacrylate) (PHEMA) cryogel. The PGMA beads were prepared by dispersion polymerization. The PGMA beads were modified with Cibacron Blue F3GA and iminodiacetic acid (IDA)-Cu 2+ for simultaneous albumin and IgG depletion, respectively. The PHEMA cryogel was synthesized by free radical polymerization in the presence of the modified PGMA beads. The PHEMA and PHEMA/PGMA composite cryogels were characterized by swelling measurements and scanning electron microscopy (SEM). Protein depletion studies were carried out in a continuous experimental set-up in a stacked column. Albumin adsorption capacity of the PGMA-Cibacron Blue F3GA beads embedded PHEMA cryogel (PHEMA/PGMA-Cibacron Blue F3GA) was 342 mg/g and IgG adsorption capacity of the PGMA-IDA-Cu 2+ beads embedded PHEMA cryogel (PHEMA/PGMA-IDA-Cu 2+ ) was 257 mg/g. The composite cryogels depleted albumin and IgG from human serum with 89.4% and 93.6% efficiency, respectively. High desorption values (over 90% for both modified cryogels) were achieved with 0.05 M phosphate buffer containing1.0 M NaCl.

N-Methacryloyl-l-phenylalanine (MAPA) containing poly(2-hydroxyethylmethacrylate) based magnetic [mag-poly(HEMA–MAPA)] nanobeads was prepared for lysozyme purification form chicken egg white. MAPA was synthesized by reacting methacryloyl chloride with l-phenylalanine methyl ester and provided hydrophobic functionality to the nanobeads. Size of mag-poly(HEMA–MAPA) nanobeads was 386 nm and obtained by surfactant free emulsion polymerization of HEMA and MAPA having a specific surface area of 580 m2/g. Mag-poly(HEMA–MAPA) nanobeads were characterized by FTIR, AFM, TEM, ESR, and elemental analysis. Lysozyme adsorption experiments were investigated under different conditions in batch system (i.e., medium pH, protein concentration, temperature, salt type). Lysozyme adsorption capacity of mag-poly(HEMA) and mag-poly(HEMA–MAPA) nanobeads from aqueous solutions was estimated as 24 and 517 mg/g, respectively. Lysozyme molecules were desorbed with 50% ethylene glycol solution with 98% recovery. It was observed that mag-poly(HEMA–MAPA) nanobeads can be used without significant decrease in lysozyme adsorption capacity after ten adsorption–desorption cycles. Mag-poly(HEMA–MAPA) nanobeads was used for the purification of lysozyme from chicken egg white. Purity of lysozyme was estimated by SDS-PAGE.

Poly(hydroxyethyl methacrylate) (PHEMA) nanobeads with an average size of 300 nm in diameter and with a polydispersity index of 1.156 were produced by a surfactant free emulsion polymerization. Specific surface area of the PHEMA nanobeads was found to be 996 m 2 /g. Imidazole containing 3-(2-imidazoline-1-yl)propyl(triethoxysilane) (IMEO) was used as a metal-chelating ligand. IMEO was covalently attached to the nanobeads. PHEMA-IMEO nanobeads were used for the removal of copper(II) ions from aqueous solutions. To evaluate the degree of IMEO loading, the PHEMA nanobeads were subjected to Si analysis by using flame atomizer atomic absorption spectrometer and it was estimated as 973 µmol IMEO/g of polymer. The PHEMA nanobeads were characterized by transmission electron microscopy and fourier transform infrared spectroscopy. Adsorption equilibrium was achieved in about 8 min. The adsorption of Cu 2+ ions onto the PHEMA nanobeads was negligible (0.2 mg/g). The IMEO attachment into the PHEMA nanobeads significantly increased the Cu 2+ adsorption capacity (58 mg/g). Adsorption capacity of the PHEMA-IMEO nanobeads increased significantly with increasing concentration. The adsorption of Cu 2+ ions increased with increasing pH and reached a plateau value at around pH 5.0. Competitive heavy metal adsorption from aqueous solutions containing Cu + , Cd 2+ , Pb 2+ and Hg 2+ was also investigated. The adsorption capacities are 61.4 mg/g (966.9 µmol/g) for Cu 2+ ; 180.5 mg/g (899.8 µmol/g) for Hg 2+ ; 34.9 mg/g (310.5 µmol/g) for Cd 2+ and 14.3 mg/g (69 µmol/g) for Pb 2+ . The affinity order in molar basis is observed as Cu 2+ > Hg 2+ > Cd 2+ > Pb 2+ . These results may be considered as an indication of higher specificity of the PHEMA-IMEO nanobeads for the Cu 2+ comparing to other ions. Consecutive adsorption and elution operations showed the feasibility of repeated use for PHEMA-IMEO nanobeads.

WOS: 000269139900058, The purification of hyaluronic acid (HA) is relatively significant to use in biomedical applications. The structure of HA is formed by the repetitive units of glucuronic acid and N-acetyl glucosamine. In this study, glucuronic acid-imprinted microbeads have been supplied for the purification of HA from cell culture (Streptococcus equi). Histidine-functional monomer. methacryloylamidohistidine (MAH) was chosen as the metal-complexing monomer. The glucuronic acid-imprinted poly(ethyleneglycoldimethacrylate-MAH-Copper(II)) [p(EDMA-MAH-Cu2+)] microbeads have been synthesized by typical suspension polymerization procedure. The template glucuronic acid has been removed by employing 5 M methanolic KOH solution. p (EDMA-MAH-Cu2+) microbeads have been characterized by elemental analysis, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) images and swelling studies. Moreover, HA adsorption experiments have been performed in a batch experimental set-up. Purification of HA from cell culture supernatant has been also investigated by determining the hyaluronidase activity using purified HA as substrate. The glucuronic acid imprinted p(EDMA-MAH-Cu2+) particles can be used many times with no significant loss in adsorption capacities. Also, the selectivity of prepared molecular imprinted polymers (MIP) has been examined. Results have showed that MIP particles are 19 times more selective for glucuronic acid than N-acetylglucose amine

WOS: 000262136700023, The aim of this study is to utilize ion-imprinted magnetic beads in the selective removal of Cd2+ ions out of human plasma overdosed with Cd2+ ions. The Cd (2+) imprinted magnetic poly(HEMA-MAC) (mPHEMAC-Cd2+) beads were produced by suspension polymerization in the presence of magnetite Fe3O4 in a nano-powder form. The template Cd2+ ions could be reversibly detached from the matrix to form mPHEMAC-Cd2+ beads using 0.1 M thiourea solution. The specific surface area of the mPHEMAC-Cd2+ beads was found to be 24.7 m(2)/g. The MAC and Fe3O4 contents of the mPHEMAC-Cd2+ beads were found to be 41.8 mu mol/g polymer and 8.2% on the average. The Cd2+ adsorption capacity of mPHEMAC-Cd2+ columns decreased drastically from 48.8 mu mol/g to 20.0 mu mol/g as the flow rate is increased from 0.50 ml/min to 3.0 ml/min. The maximum adsorption capacity of the mPHEMAC-Cd2+ beads was determined to be 48.8 mu mol Cd2+/g on the average. The relative selectivity coefficients of the mPHEMAC beads for Cd2+/Pb2+ and Cd2+/Zn2+ were 22.6 and 160.7 times greater than those of the non-imprinted magnetic PHEMAC (mPHEMAC) beads, respectively. The mPHEMAC-Cd2+ beads are reusable for many times with no significant decrease in their adsorption capacities

WOS: 000243833600018, Molecular recognition-based separation techniques have received much attention in chemistry and biology because of their high selectivity for target molecules. The aim of this study is to prepare lysozyme-imprinted polymers which can be used for the purification of lysozyme from aqueous solutions and egg white. N-methacryloyl-(L)-histidinemethylester (MAH) was chosen as the metal-complexing monomer. In the first step, Cu2+ was complexed with MAH and the lysozyme-imprinted poly(HEMA-MAH) [Lys-MIP] particles were synthesized by UV-initiated bulk polymerization. After that, the template (i.e., lysozyme) were removed using 0.1 M NaCl solution. The specific surface area of the Lys-MIP particles was found to be 22.9 m(2)/g with a size range of 20-63 mu m in diameter and the swelling ratio was 57%. According to the elemental analysis results, the particles contained 421 mu mol MAH/g polymer. The maximum lysozyme adsorption capacity was 12.1 mg/g polymer. The relative selectivity coefficients of imprinted particles for lysozyme/human serum albumin and lysozyme/cytochrome c were 3.6 and 4.1 times greater than NIP particles, respectively. Purification of lysozyme from egg-white was also monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purity of the desorbed lysozyme was about 89% with recovery about 84%. The Lys-MIP particles could be used many times without decreasing their adsorption capacities significantly

N-methacryloyl-(l)-glutamic acid (MAGA) was synthesized using methacryloyl chloride and l-glutamic acid methyl ester as a metal-complexing ligand and/or comonomer. MAGA was characterized by FTIR and NMR. Spherical beads with an average diameter of 150–200 μm were obtained by suspension polymerization of MAGA and 2-hydroxyethyl methacrylate (HEMA) performed in an aqueous dispersion medium. Poly(HEMA-MAGA) beads were characterized by swelling studies, surface area measurements and elemental analysis. Poly(HEMA-MAGA) beads have a specific surface area of 56.7 m2/g. Poly(HEMA-MAGA) beads were used in the removal studies of Pb2+ ions. Adsorption equilibrium was achieved in about 60 min. The adsorption of Pb2+ ions onto PHEMA beads was negligible (0.38 mg/g). The MAGA incorporation into the polymer structure significantly increased the lead adsorption capacity (348 mg/g). The adsorption of Pb2+ ions increased with increasing pH and reached a plateau value at around pH 5.0. Competitive adsorption of heavy metal ions from synthetic wastewater was also studied. The adsorption capacities are 42.5 mg/g for Pb2+, 26.8 mg/g for Hg2+ and 17.6 mg/g for Cd2+ at 0.5 mmol/l metal concentration. Consecutive adsorption and elution operations showed the feasibility of repeated use for poly(HEMA-MAGA) chelating beads.