Adsorption of IgG on spacer‐arm and L‐arginine ligand attached poly(GMA/MMA/EGDMA) beads

This work presents data on human immunoglobulin G (HIgG) adsorption onto L ‐arginine ligand attached poly(GMA/MMA/EGDMA)‐based affinity beads which were synthesized from methyl methacrylate (MMA) and glycidiyl methacrylate (GMA) in the presence of a crosslinker (i.e., ethylene glycol dimethacrylate; EGDMA) by suspension polymerization. The epoxy groups of the poly(GMA/MMA/EGDMA) beads were converted into amino groups after reaction with ammonia or 1,6‐diaminohexane (i.e., spacer‐arm). With L ‐arginine as a ligand, it was covalently immobilized on the aminated (poly(GMA/MMA/EGDMA)‐ AA) and/or the spacer‐arm attached (poly(GMA/MMA/EGDMA)‐SA) beads, using glutaric dialdehyde as a coupling agent. Both affinity poly(GMA/MMA/EGDMA)‐based beads were used in HIgG adsorption/desorption studies under defined pH, ionic strength, or temperature conditions in a batch reactor, using acid‐treated poly(GMA/MMA/EGDMA) beads as a control system. The poly(GMA/MMA/EGDMA)‐SA affinity beads resulted in an increase in the adsorption capacity to HIgG compared with the aminated counterpart (i.e., poly(GMA/MMA/EGDMA)‐AA). The maximum adsorption capacities of the poly(GMA/MMA/EGDMA)‐AA and poly(GMA/MMA/EGDMA)‐SA affinity beads were found to be 112.36 and 142 mg g^?1, and the affinity constants (K_d), evaluated by the Langmuir model, were 2.48 × 10^?7 and 6.98 × 10^?7M, respectively. Adsorption capacities of the poly(GMA/MMA/EGDMA)‐AA and poly(GMA/MMA/EGDMA)‐SA were decreased with HIgG by increasing the ionic strength adjusted with NaCl. Adsorption kinetic of HIgG onto both affinity adsorbents was analyzed with first‐ and second‐order kinetic equations. The first‐order equation fitted well with the experimental data. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 104: 672–679, 2007     

Human serum albumin adsorption on poly[(glycidyl methacrylate)‐co‐(methyl methacrylate)] beads modified with a spacer‐arm‐attached L‐histidine ligand

Poly(GMA/MMA) beads were synthesized from glycidyl methacrylate (GMA) and methyl methacrylate (MMA) in the presence of a cross‐linker (i.e. ethyleneglycol dimethacrylate) (EGDMA) via suspension polymerization. The epoxy groups of the poly(GMA/MMA) beads were converted into amino groups with either ammonia or 1,6‐diaminohexane (i.e. spacer‐arm). An L ‐histidine ligand was then covalently immobilized on the aminated (poly(GMA/MMA)‐AH) and/or the spacer‐arm attached (poly(GMA/MMA)‐SAH) beads using glutaric dialdehyde as a coupling agent. Both affinity adsorbents were used in human serum albumin (HSA) adsorption/desorption studies under defined pH, ionic strength or temperature conditions in a batch reactor. The spacer‐arm attached affinity adsorbent resulted in an increase in the adsorption capacity to HSA when compared to the aminated counterpart (i.e. poly(GMA/MMA)‐AH). The maximum adsorption capacities of the affinity adsorbents were found to be significantly high, i.e. 43.7 and 80.2 mg g^?1 (of the beads), while the affinity constants, evaluated by the Langmuir model, were 3.96 × 10^?7 and 9.53 × 10^?7 mol L^?1 for poly(GMA/MMA)‐AH and poly(GMA/MMA)‐SAH, respectively. The adsorption capacities of the affinity adsorbents were decreased for HSA by increasing the ionic strength, adjusted with NaCl. The adsorption kinetics of HSA were analysed by using pseudo‐first and pseudo‐second‐order equations. The second‐order equation fitted well with the experimental data. Copyright © 2005 Society of Chemical Industry     

Vinyl triazole carrying metal‐chelated beads for the reversible immobilization of glucoamylase

Poly(ethylene glycol dimethacrylate–1‐vinyl‐1,2,4‐triazole) [poly(EGDMA–VTAZ)] beads with an average diameter of 100–200 μm were obtained by the copolymerization of ethylene glycol dimethacrylate (EGDMA) with 1‐vinyl‐1,2,4‐triazole (VTAZ). The copolymer hydrogel bead composition was determined by elemental analysis and was found to contain 5 EGDMA monomer units for each VTAZ monomer unit. The poly(EGDMA–VTAZ) beads were characterized by swelling studies and scanning electron microscopy (SEM). The specific surface area of the poly(EGDMA–VTAZ) beads was found 65.8 m²/g. Cu²⁺ ions were chelated on the poly(EGDMA–VTAZ) beads. The Cu²⁺ loading was 82.6 μmol/g of support. Cu²⁺‐chelated poly(EGDMA–VTAZ) beads with a swelling ratio of 84% were used in the immobilization of Aspergillus niger glucoamylase in a batch system. The maximum glucoamylase adsorption capacity of the poly(EGDMA–VTAZ)–Cu²⁺ beads was 104 mg/g at pH 6.5. The adsorption isotherm of the poly(EGDMA–VTAZ)–Cu²⁺ beads fitted well with the Langmuir model. Adsorption kinetics data were tested with pseudo‐first‐ and second‐order models. The kinetic studies showed that the adsorption followed a pseudo‐second‐order reaction model. The Michaelis constant value for the immobilized glucoamylase (1.15 mg/mL) was higher than that for free glucoamylase (1.00 mg/mL). The maximum initial rate of the reaction values were 42.9 U/mg for the free enzyme and 33.3 U/mg for the immobilized enzyme. The optimum temperature for the immobilized preparation of poly(EGDMA–VTAZ)–Cu²⁺–glucoamylase was 65°C; this was 5°C higher than that of the free enzyme at 60°C. The glucoamylase adsorption capacity and adsorbed enzyme activity slightly decreased after 10 batch successive reactions; this demonstrated the usefulness of the enzyme‐loaded beads in biocatalytic applications. The storage stability was found to increase with immobilization. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011     

Covalent immobilization of β‐galactosidase onto electrospun nanofibers of poly (AN‐co‐MMA) copolymer

Immobilization of β‐galactosidase in poly (acrylonitrile‐co‐methyl methacrylate) poly (AN‐co‐MMA) Nanofibers was studied by electrospinning, and a spacer‐arm i.e., (Polyethyleneimine (PEI)) was covalently attached by the reaction of carbonyl groups of Poly (AN‐co‐MMA) nanofibers. β‐galactosidase was then covalently immobilized through the spacer‐arm of the Poly (AN‐co‐MMA) nanofibers by using glutaraldehyde (GA) as a coupling agent. Nanofibers mode of interaction was proven by FTIR and thermal gravimetric analysis and supported by morphological changes recognized through SEM examination. Factors affecting the modification process such as PEI concentration, reaction time, and reaction temperature have been studied. Its influence on the amount of coupled PEI was consequently correlated to the changes of the catalytic activity and the retained activity of immobilized enzyme, the main parameters judging the success of the immobilization process. Evidences of Poly (AN‐co‐MMA) nanofibers modification were extracted from morphological changes recognized through SEM examination. The maximum activity (V_max) and michaelis constant (K_m) of immobilized enzyme were found to be 8.8 μmole/min mg protein and 236.7 mM, respectively. Stabilities of the immobilized β‐galactosidase were obviously improved. The optimum temperature for β‐galactosidase immobilized on the spacer‐arm attached nanofiber was 5°C higher than that of the free enzyme and was also significantly broader. The immobilized β‐galactosidase had better resistance to temperature inactivation than did the free form. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Synthesis and characterization of a porous poly(hydroxyethylmethacrylate‐co‐ethylene glycol dimethacrylate)‐based hydrogel device for the implantable delivery of insulin

Poly(hydroxyethylmethacrylate‐co‐ethylene glycol dimethacrylate) [poly(HEMA‐co‐EGDMA)]‐based hydrogel devices were synthesized by a free‐radical polymerization reaction with 2‐hydroxyethylmethacrylate as the monomer, different concentrations of ethylene glycol dimethacrylate (EGDMA) as the crosslinking agent, and ammonium persulfate/N,N,N′,N′‐tetra‐methyl ethylenediamine as the free‐radical initiator. The porosity of the poly(HEMA‐co‐EGDMA) hydrogels was controlled with water as the porogen. The Fourier transform infrared spectrum of poly(HEMA‐co‐EGDMA) showed absorption bands associated with ? C?O stretching at 1714 cm^?1, C? O? C stretching vibrations at 1152 cm^?1, and a broad band at 3500–3800 cm^?1 corresponding to ? OH stretching. Atomic force microscopy studies showed that the hydrogel containing 67% water had pores in the range of 3500–9000 nm, whereas the hydrogel containing 7% water did not show measurable pores. The hydrogel synthesized with 1% EGDMA showed 50% thallium‐201 release within the first 30 min and about 80% release within 60 min. In vitro insulin‐release studies suggested that the hydrogel with 27% water showed sustained release up to 120 min, whereas the hydrogels with 47 and 67% water showed that nearly all of the insulin was released within 60 min. Hydrogel devices synthesized with 27% water and filled with insulin particles showed sustained release for up to 8 days, whereas the hydrogels synthesized with 47 and 67% water released insulin completely within 3 days of administration. Animal studies suggested that the hydrogel devices synthesized with 27% water and filled with insulin‐loaded particles (120 IU) were able to control blood glucose levels for up to 5 days after implantation. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

Reactive red 120 and NI(II) derived poly(2‐hydroxyethyl methacrylate) nanoparticles for urease adsorption

Non‐porous poly(2‐hydroxyethyl methacrylate) [p(HEMA)] nanoparticles were prepared by surfactant free emulsion polymerization. The p(HEMA) nanoparticles was about 200 nm diameter, spherical form, and non‐porous. Reactive Red 120 (RR 120) was covalently attached to the p(HEMA) nanoparticles and Ni(II) ions were incorporated to attach dye molecules. Urease was immobilized onto RR120‐Ni(II) attached p(HEMA) nanoparticles via adsorption. The maximum urease adsorption capacity of RR120‐Ni(II) attached p(HEMA) nanoparticles was 480.01 mg g^?1 nanoparticles at pH 7.0 in phosphate buffer. It was observed that urease could be repeatedly adsorbed and desorbed without significant loss in adsorption amount. K_m values were 21.50 and 34.06 mM for the free and adsorbed enzyme. The V_max values were 4 U for the free enzyme and 3.3 U for the adsorbed enzyme. The optimum pH was 25 mM pH 7 phosphate buffer for free and adsorbed enzyme. The optimum temperature was determined at 35°C and 55°C for the free and adsorbed enzyme, respectively. These findings show considerable promise for this material as an adsorption matrix in biotechnological applications. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39757.     

Heparin‐immobilized poly(2‐hydroxyethylmethacrylate)‐based microspheres

Poly(2‐hydroxyethylmethacrylate) (PHEMA)‐based microspheres (150–200 µm in diameter) were produced by a modified suspension polymerization of different type of comonomers—namely, acrylic acid, dimethylaminoethyl‐methacrylate, and methylmethacrylate. These microspheres were activated with cyanogen bromide (CNBr) at pH 11.5, and heparin molecules were then immobilized through covalent bonds. The amount of immobilized heparin was controlled by changing the initial concentration of CNBr and heparin. The increase in the initial concentrations of both CNBr and heparin caused an increase in the amount of heparin immobilized onto microspheres for all polymer surfaces. The maximum heparin immobilization was observed on the PHEMA homopolymer microspheres (180 mg/g). The plain and heparin‐immobilized microspheres were contacted with blood in in vitro systems and in ex vivo animal experiments. Loss of the blood cells and clotting times were followed. Anticoagulant effect of the immobilized heparin was clearly observed with blood coagulation experiments. Loss of cells in the blood contacting with heparin‐immobilized microspheres was significantly lower than those observed with the plain microspheres. Bovine serum albumin adsorption onto the microspheres containing heparin on their surfaces was also studied. High albumin adsorption values (up to 127 mg/g) were observed in which the heparin‐immobilized PHEMA microspheres were used. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 74: 655–662, 1999     

Congo red attached poly(EGDMA–HEMA) microspheres as specific sorbents for removal of cadmium ions

Poly[ethyleneglycoldimethacrylate (EGDMA)–hydroxyethylmethacrylate (HEMA)] microspheres (150–200 μm in diameter) were produced by suspension copolymerization of EGDMA and HEMA in an aqueous medium. Toluene was included in the formulations in order to produce water-swellable microspheres. Poly(vinyl alcohol) and benzoyl peroxide were used as stabilizer and initiator, respectively. Congo red was chemically attached to the microspheres as a metal chelating ligand for specific adsorption of heavy metal ions. These sorbents were characterized by an optical microscopy and a FTIR. Adsorption/desorption of cadmium (Cd²⁺) ions from aqueous solutions on these sorbents were investigated in batch equilibrium experiments by using an atomic absorption spectroscopy with a graphite furnace atomizer. The maximum cadmium adsorption on to the dye-attached microspheres (i.e., by complex formation) was about 18.3 mg Cd²⁺ ions/g polymer, which was observed at pH 6.8. While adsorption onto the plain poly(EGDMA–HEMA) microspheres (i.e., nonspecific adsorption) was about 0.93 mg Cd²⁺ ions/g polymer at the same conditions. More than 90% of the adsorbed cadmium was desorbed in 1 h by using 2M NaCl as an eluant. The resorption capacity of the sorbent did not significantly decrease during repeated sorption–desorption cycling. © 1996 John Wiley & Sons, Inc.     

Porous dye affinity beads for albumin separation from human plasma

Porous polymeric beads were obtained by the suspension polymerization of 2‐hydroxyethyl methacrylate (HEMA) and ethylene glycol dimethacrylate (EGDMA). Poly(HEMA–EGDMA) beads were characterized by surfacearea measurements, swelling studies, FTIR, scanning electron microscopy (SEM), and elemental analysis. Poly (HEMA–EGDMA) beads had a specific surface area of 56 m²/g. SEM observations showed that the poly(HEMA–EGDMA) beads abounded macropores. Poly(HEMA–EGDMA) beads with a swelling ratio of 55%, and containing different amounts of Reactive Red 120 (9.2–39.8 μmol/g) were used in the adsorption/desorption of human serum albumin (HSA) from aqueous solutions and human plasma. The nonspecific adsorption of HSA was very low (0.2 mg/g). The maximum HSA adsorption amount from aqueous solution in phosphate buffer was 60.1 mg/g at pH 5.0. Higher HSA adsorption value was obtained from human plasma (up to 95.7 mg/g) with a purity of 88%. The equilibrium monolayer adsorption amount, Q_max was determined as 172.4 mg/g. The dimensionless separation factor (R_L) value shows that the adsorption behavior of HSA onto the Reactive Red 120 attached poly(HEMA–EGDMA) beads was favorable (0 < R_L < 1). Desorption of HSA from Reactive Red 120 attached poly (HEMA–EGDMA) beads was performed using 0.1M Tris/HCl buffer containing 0.5M NaCl. It was observed that HSA could be repeatedly adsorbed and desorbed with Reactive Red 120‐attached poly(HEMA–EGDMA) beads without significant loss in the adsorption amount. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci, 2007     

Invertase immobilized on spacer‐arm attached poly(hydroxyethyl methacrylate) membrane: Preparation and properties

Microporous poly(2‐hydroxyethyl methacrylate) (pHEMA) membrane was prepared by UV‐initiated photopolymerization. The spacer arm (i.e., hexamethylene diamine) was attached covalently and then invertase was immobilized by the condensation reaction of the amino groups of the spacer arm with carboxyl groups of the enzyme in the presence of carbodiimides. The values of the Michael's constant K_m of invertase were significantly larger (ca. 2.5 times) upon immobilization, indicating decreased affinity by the enzyme for its substrate, whereas V_max was smaller for the immobilized invertase. Immobilization improved the pH stability of the enzyme as well as its temperature stability. Thermal stability was found to increase with immobilization and at 70°C the half times for the activity decay were 12 min for the free enzyme and 41 min for the immobilized enzyme. The immobilized enzyme activity was found to be quite stable in repeated experiments. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 75: 1685–1692, 2000     

New generation polymeric nanospheres for catalase immobilization

Monosize poly(2‐hydroxyethyl methacrylate‐co‐N‐methacryloly‐L ‐histidinemethylester) [mon‐poly(HEMA‐MAH)] nanospheres were prepared via surfactant‐free emulsion polymerization method. L ‐Histidine groups of the mon‐poly(HEMA‐MAH) nanospheres were chelated with Fe³⁺ ions. Mon‐poly(HEMA‐MAH) nanospheres were characterized by Fourier transform infrared spectroscopy, proton NMR, and scanning electron microscopy. Particle size of the mon‐poly(HEMA‐MAH) nanospheres was measured by Zeta Sizer. Elemental analysis of MAH for nitrogen was estimated as 0.94 mmol/g polymer. The catalase immobilized onto the mon‐poly(HEMA‐MAH)–Fe³⁺ nanospheres resulted in increasing the enzyme stability with time. Optimum operational temperature for both immobilized preparations was the same, and the temperature profiles of the immobilized preparations were significantly broader. It was observed that enzyme could be repeatedly adsorbed and desorbed on the mon‐poly(HEMA‐MAH)–Fe³⁺ nanospheres without loss of adsorption capacity or enzymic activity. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Covalent immobilization of trypsin onto thermo‐sensitive poly(N‐isopropylacrylamide‐co‐acrylic acid) microspheres with high activity and stability

Poly(N‐isopropylacrylamide‐co‐acrylic acid) (P(NIPAM‐co‐AA)) microspheres with a high copolymerized AA content were fabricated using rapid membrane emulsification technique. The uniform size, good hydrophilicity, and thermo sensitivity of the microspheres were favorable for trypsin immobilization. Trypsin molecules were immobilized onto the microspheres surfaces by covalent attachment. The effects of various parameters such as immobilization pH value, enzyme concentration, concentration of buffer solution, and immobilization time on protein loading amount and enzyme activity were systematically investigated. Under the optimum conditions, the protein loading was 493 ± 20 mg g^?1 and the activity yield of immobilized trypsin was 155% ± 3%. The maximum activity (V_max) and Michaelis constant (K_m) of immobilized enzyme were found to be 0.74 μM s^?1 and 0.54 mM, respectively. The immobilized trypsin showed better thermal and storage stability than the free trypsin. The enzyme‐immobilized microspheres with high protein loading amount still can show a thermo reversible phase transition behavior. The research could provide a strategy to immobilize enzyme for application in proteomics. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43343.     

DNA adsorption on a poly‐L‐lysine‐immobilized poly(2‐hydroxyethyl methacrylate) membrane

The DNA adsorption properties of poly‐L ‐lysine‐immobilized poly(2‐hydroxyethyl methacrylate) (pHEMA) membrane were investigated. The pHEMA membrane was prepared by UV‐initiated photopolymerization and activated with epichlorohydrin. Poly‐L ‐lysine was then immobilized on the activated pHEMA membrane by covalent bonding, via a direct chemical reaction between the amino group of poly‐L ‐lysine and the epoxy group of pHEMA. The poly‐L ‐lysine content of the membrane was determined as 1537 mg m^?2. The poly‐L ‐lysine‐immobilized membrane was utilized as an adsorbent in DNA adsorption experiments. The maximum adsorption of DNA on the poly‐L ‐lysine‐immobilized pHEMA membrane was observed at 4 °C from phosphate‐buffered salt solution (pH 7.4, 0.1 M; NaCl 0.5 M) containing different amounts of DNA. The non‐specific adsorption of DNA on the plain pHEMA membrane was low (about 263 mg m^?2). Higher DNA adsorption values (up to 5849 mg m^?2) were obtained in which the poly‐L ‐lysine‐immobilized pHEMA membrane was used. Copyright © 2003 Society of Chemical Industry     

Synthesis,characterization, and in vitro release of ibuprofen from poly(MMA‐HEMA) copolymeric core–shell hydrogel microspheres for biomedical applications

This article describes the development of a new crosslinked poly(methyl methacrylate‐2‐hydroxyethyl methacrylate) copolymeric core–shell hydrogel microsphere incorporated with ibuprofen for potential applications in bone implants. Initially poly(methyl methacrylate) (PMMA) core microspheres were prepared by free‐radical initiation technique. On these core microspheres, 2‐hydroxyethyl methacrylate (HEMA) was polymerized by swelling PMMA microspheres with the HEMA monomer by using ascorbic acid and ammonium persulfate. Crosslinking monomers such as ethylene glycol dimethacrylate (EGDMA) has also been included along with HEMA for polymerization. By this technique, it was possible to obtain core–shell‐type microspheres. The core is a hard PMMA microsphere having a hydrophilic poly(HEMA) shell coat on it. These microspheres are highly hydrophilic as compared to PMMA microspheres. The size of the hydrogel microspheres almost doubled when swollen in benzyl alcohol. These microspheres were characterized by various techniques such as optical microscopy, scanning electron microscopy, Fourier‐transformed infrared spectroscopy, thermogravimetric analysis, and differential scanning calorimetry. The particle size of both microspheres was analyzed by using Malvern Master Sizer/E particle size analyzer. The in vitro release of ibuprofen from both microspheres showed near zero‐order patterns. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 83: 3045–3054, 2002; DOI 10.1002/app.10310     

Metal chelating properties of Cibacron Blue F3GA‐derived poly(EGDMA‐HEMA) microbeads

Metal chelating properties of Cibacron Blue F3GA‐derived poly(EGDMA‐HEMA) microbeads have been studied. Poly(EGDMA‐HEMA) microbeads were prepared by suspension copolymerization of ethylene glycol dimethacrylate (EGDMA) and hydroxy‐ethyl methacrylate (HEMA) by using poly(vinyl alcohol), benzoyl peroxide, and toluene as the stabilizer, the initiator, and the pore‐former, respectively. Cibacron Blue F3GA was covalently attached to the microbeads via the nucleophilic substitution reaction between the chloride of its triazine ring and the hydroxyl groups of the HEMA, under alkaline conditions. Microbeads (150–200 μm in diameter) with a swelling ratio of 55%, and carrying 16.5 μmol Cibacron Blue F3GA/g polymer were used in the adsorption/desorption studies. Adsorption capacity of the microbeads for the selected metal ions, i.e., Cu(II), Zn(II), Cd(II), Fe(III), and Pb(II) were investigated in aqueous media containing different amounts of these ions (5–200 ppm) and at different pH values (2.0–7.0). The maximum adsorptions of metal ions onto the Cibacron Blue F3GA‐derived microbeads were 0.19 mmol/g for Cu(II), 0.34 mmol/g for Zn(II), 0.40 mmol/g for Cd(II), 0.91 mmol/g for Fe(III), and 1.05 mmol/g for Pb(II). Desorption of metal ions were studied by using 0.1 M HNO₃. High desorption ratios (up to 97%) were observed in all cases. Repeated adsorption/desorption operations showed the feasibility of repeated use of this novel sorbent system. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 71: 1397–1403, 1999     

Chitosan‐grafted poly(hydroxyethyl methacrylate‐co‐glycidyl methacrylate) membranes for reversible enzyme immobilization

Epoxy group‐containing poly(hydroxyethyl methacrylate/glycidyl methacrylate), p(HEMA/GMA), membrane was prepared by UV initiated photopolymerization. The membrane was grafted with chitosan (CH) and some of them were chelated with Fe(III) ions. The CH grafted, p(HEMA/GMA), and Fe(III) ions incorporated p(HEMA/GMA)‐CH‐Fe(III) membranes were used for glucose oxidase (GOD) immobilization via adsorption. The maximum enzyme immobilization capacity of the p(HEMA/GMA)‐CH and p(HEMA/GMA)‐CH‐Fe(III) membranes were 0.89 and 1.36 mg/mL, respectively. The optimal pH value for the immobilized GOD preparations is found to have shifted 0.5 units to more acidic pH 5.0. Optimum temperature for both immobilized preparations was 10°C higher than that of the free enzyme and was significantly broader at higher temperatures. The apparent K_m values were found to be 6.9 and 5.8 mM for the adsorbed GOD on p(HEMA/GMA)‐CH and p(HEMA/GMA)‐CH‐Fe(III) membranes, respectively. In addition, all the membranes surfaces were characterized by contact angle measurements. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 3084–3093, 2007     

Nonporous monosize polymeric sorbents: Dye and metal chelate affinity separation of lysozyme

Lysozyme adsorption onto dye‐attached nonporous monosize poly(2‐hydroxyethyl‐methacrylate‐methylmethacrylate) [poly(HEMA‐MMA)] microspheres was investigated. Poly(HEMA‐MMA) microspheres were prepared by dispersion polymerization. The monochloro‐triazine dye, Cibacron Blue F3GA, was immobilized covalently as dye–ligand. These dye‐affinity microspheres were used in the lysozyme adsorption–desorption studies. The effect of initial concentration of lysozyme and medium pH on the adsorption efficiency of dye‐attached and metal‐chelated microspheres were studied in a batch reactor. Effect of Cu(II) chelation on lysozyme adsorption was also studied. The nonspecific adsorption of lysozyme on the poly(HEMA‐MMA) microspheres was 3.6 mg/g. Cibacron Blue F3GA attachment significantly increased the lysozyme adsorption up to 247.8 mg/g. Lysozyme adsorption capacity of the Cu(II) incorporated microspheres (318.9 mg/g) was greater than that of the Cibacron Blue F3GA‐attached microspheres. Significant amount of the adsorbed lysozyme (up to 97%) was desorbed in 1 h in the desorption medium containing 1.0M NaSCN at pH 8.0 and 25 mM EDTA at pH 4.9. In order to examine the effects of separation conditions on possible conformational changes of lysozyme structure, fluorescence spectrophotometry was employed. We conclude that dye‐ and metal‐chelate affinity chromatography with poly(HEMA‐MMA) microspheres can be applied for lysozyme separation without causing any significant changes and denaturation. Repeated adsorption/desorption processes showed that these novel dye‐attached monosize microspheres are suitable for lysozyme adsorption. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 76: 115–124, 2000     

Synthesis and characterization of poly(HEMA‐co‐MMA)‐g‐POSS nanocomposites by combination of reversible addition fragmentation chain transfer polymerization and click chemistry

New hybrid poly(hydroxyethyl methacrylate‐co‐methyl methacrylate)‐g‐polyhedral oligosilsesquioxane [poly(HEMA‐co‐MMA)‐g‐POSS] nanocomposites were synthesized by the combination of reversible addition fragmentation chain transfer (RAFT) polymerization and click chemistry using a grafting to protocol. Initially, the random copolymer poly(HEMA‐co‐MMA) was prepared by RAFT polymerization of HEMA and MMA. Alkynyl side groups were introduced onto the polymeric backbones by esterification reaction between 4‐pentynoic acid and the hydroxyl groups on poly(HEMA‐co‐MMA). Azide‐substituted POSS (POSS? N₃) was prepared by the reaction of chloropropyl‐heptaisobutyl‐substituted POSS with NaN₃. The click reaction of poly(HEMA‐co‐MMA)‐alkyne and POSS? N₃ using CuBr/PMDEATA as a catalyst afforded poly(HEMA‐co‐MMA)‐g‐POSS. The structure of the organic/inorganic hybrid material was investigated by Fourier transformed infrared, ¹H‐NMR, and ²⁹Si‐NMR. The elemental mapping analysis of the hybrid using X‐ray photoelectron spectroscopy and EDX also suggest the formation of poly(HEMA‐co‐MMA)‐anchored POSS nanocomposites. The XRD spectrum of the nanocomposites gives evidence that the incorporation of POSS moiety leads to a hybrid physical structure. The morphological feature of the hybrid nanocomposites as captured by field emission scanning electron microscopy and transmission electron microscopic analyses indicate that a thick layer of polymer brushes was immobilized on the POSS cubic nanostructures. The gel permeation chromatography analysis of poly(HEMA‐co‐MMA) and poly(HEMA‐co‐MMA)‐g‐POSS further suggests the preparation of nanocomposites by the combination of RAFT and click chemistry. The thermogravimetric analysis revealed that the thermal property of the poly(HEMA‐co‐MMA) copolymer was significantly improved by the inclusion of POSS in the copolymer matrix. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Adsorption of bilirubin on poly‐L‐lysine‐containing nylon membranes: applications in affinity chromatography

Microporous polyamide membranes were activated by 1,1′‐carbonyldiimidazole (CDI) and subsequently bound with hydroxyethyl cellulose (HEC) to amplify reactive groups. Then poly‐L ‐lysine (PLL) as ligand was immobilized onto the HEC‐nylon membranes. The contents in HEC and PLL of PLL‐attached membranes were 153.2 and 63.8 mg (g nylon membrane)^?1, respectively. Such PLL‐HEC affinity membranes were used to adsorb bilirubin from bilirubin‐phosphate and bilirubin‐albumin solutions. The adsorption mechanism of bilirubin and the effects of temperature and ionic strength on adsorption were investigated by batch experiments. The results showed that the adsorption capacity increased with increasing temperature but decreased with increasing NaCl concentration, and the adsorption isotherm fitted the Freundlich model well. Dynamic experiments showed that PLL‐attached membranes can readily remove the bilirubin from bilirubin‐albumin solutions. Copyright © 2005 Society of Chemical Industry     

Preparation of narrow‐disperse or monodisperse poly{[poly(ethylene glycol) methyl ether acrylate]‐co‐(acrylic acid)} microspheres with ethyleneglycol dimethacrylate as crosslinker by distillation precipitation polymerization

Narrow‐disperse or monodisperse poly{[poly(ethylene glycol) methyl ether acrylate]‐co‐(acrylic acid)} (poly(PEGMA‐co‐AA)) microspheres were prepared by distillation precipitation polymerization with ethyleneglycol dimethacrylate (EGDMA) as crosslinker with 2,2′‐azobisisobutyronitrile as initiator in neat acetonitrile in the absence of any stabilizer, without stirring. The diameters of the resultant poly(PEGMA‐co‐AA‐co‐EGDMA) microspheres were in the range 200–700 nm with a polydispersity index of 1.01–1.14, which depended on the comonomer feed of the polymerization. The addition of the hydrogen bonding monomer acrylic acid played an essential role in the formation of narrow‐disperse or monodisperse polymer microspheres during the polymerization. Copyright © 2006 Society of Chemical Industry