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     

Glutamic acid containing supermacroporous poly(hydroxyethyl methacrylate) cryogel disks for UO22+ removal

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 m²/g. The equilibrium swelling ratio of the cryogel is 9.68 g H₂O/g for poly(HEMA–MAGA) and 8.56 g H₂O/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 UO₂²⁺ from aqueous solutions. Adsorption equilibrium of UO₂²⁺ was obtained in about 30 min. The adsorption of UO₂²⁺ ions onto the PHEMA cryogel disks was negligible (0.78 mg/g). The MAGA incorporation significantly increased the UO₂²⁺ adsorption capacity (92.5 mg/g). The adsorption process is found to be a function of pH of the UO₂²⁺ 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.     

Hydrophobic microbeads as an alternative pseudo-affinity adsorbent for recombinant human interferon-α via hydrophobic interactions

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-α (rHuIFN-α) 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 m²/g and 47.3 Å for poly(HEMA) microbeads and 18.7 m²/g and 49.8 Å for poly(HEMA–MAPA) microbeads. Adsorption experiments were performed under different conditions. Maximum rHuIFN-α 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 μm and containing 327 μmol MAPA/g microbeads. Compared with poly(HEMA–MAPA) microbeads, nonspecific rHuIFN-α 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-α structure, fluorescence spectrophotometry was employed. Repeated adsorption–elution processes showed that these microbeads are suitable for repeatable rHuIFN-α adsorption.     

Selective removal of 17β-estradiol with molecularly imprinted particle-embedded cryogel systems

The selective removal of 17β-estradiol (E2) was investigated by using molecularly E2 imprinted (MIP) particle embedded poly(hydroxyethyl methacrylate) (PHEMA) cryogel. PHEMA/MIP composite cryogel was characterized by FTIR, SEM, swelling studies, and surface area measurements. E2 adsorption studies were performed by using aqueous solutions which contain various amounts of E2. The specificity of PHEMA/MIP cryogel to recognition of E2 was performed by using cholesterol and stigmasterol. PHEMA/MIP cryogel exhibited a high binding capacity (5.32 mg/gpolymer) and high selectivity for E2 in the presence of competitive molecules, cholesterol (k(E2/cholesterol) = 7.6) and stigmasterol (k(E2/Stigmasterol) = 85.8). There is no significant decrease in adsorption capacity after several adsorption-desorption cycles.     

Chiral recognition of proteins having L-histidine residues on the surface with lanthanide ion complex incorporated-molecularly imprinted fluorescent nanoparticles

In this study, lanthanide ion complex incorporated molecularly imprinted fluorescent nanoparticles were synthesized. A combination of three novel approaches was applied for the purpose. First, lanthanide ions [Terbium(III)] were complexed with N-methacryloyl-L-histidine (MAH), polymerizable derivative of L-histidine amino acid, in order to incorporate the complex directly into the polymeric backbone. At the second stage, L-histidine molecules imprinted nanoparticles were utilized instead of whole protein imprinting in order to avoid whole drawbacks such as fragility, complexity, denaturation tendency, and conformation dependency. At the third stage following the first two steps mentioned above, imprinted L-histidine was coordinated with cupric ions [Cu(II)] to conduct the study under mild conditions. Then, molecularly imprinted fluorescent nanoparticles synthesized were used for L-histidine adsorption from aqueous solution to optimize conditions for adsorption and fluorimetric detection. Finally, usability of nanoparticles was investigated for chiral biorecognition using stereoisomer, D-histidine, racemic mixture, D,L-histidine, proteins with surface L-histidine residue, lysozyme, cytochrome C, or without ribonuclease A. The results revealed that the proposed polymerization strategy could make significant contribution to the solution of chronic problems of fluorescent component introduction into polymers. Additionally, the fluorescent nanoparticles reported here could be used for selective separation and fluorescent monitoring purposes.     

L-histidine imprinted synthetic receptor for biochromatography applications

We have proposed novel surface-imprinted beads for selective separation of cytochrome c (cyt c) by N-methacryloyl-(L)-histidine-copper(II) [MAH-Cu(II)] as a new metal-chelating monomer via metal coordination interactions and histidine template. We have combined molecular imprinting with the ability of histidine to chelate metal ions to create ligand exchange beads suitable for the binding of cyt c (surface histidine exposed protein). The histidine imprinted beads were produced by suspension polymerization of MAH-Cu(II)-L-histidine and ethylene glycol dimethacrylate. After polymerization, the template (L-histidine) was removed from the beads using methanolic KOH, thus getting histidine imprinted metal-chelate beads. L-Histidine imprinted metal-chelate beads can be used several times without considerable loss of cyt c adsorption capacity. The association constant (Ka) for the specific interaction between the template imprinted polymer and the template (L-histidine) itself were determined by Scatchard plots using L-histidine imprinted beads and found as 58,300 M(-1). Finally, we have used these histidine imprinted beads for cyt c and ribonuclease A (surface histidine exposed proteins) and enantiometric separation of D- and L-histidine by FPLC.     

Poly(glycidyl methacrylate) beads embedded cryogels for pseudo-specific affinity depletion of albumin and immunoglobulin G

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)-Cu²⁺ 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)-Cu²⁺ 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-Cu²⁺ beads embedded PHEMA cryogel (PHEMA/PGMA-IDA-Cu²⁺) 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.     

Poly(hydroxyethyl methacrylate-co-methacryloylamidotryptophane) nanospheres and their utilization as affinity adsorbents for porcine pancreas lipase adsorption

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 K_m 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.     

Preparation of cibacron blue F3GA‐attached polyamide hollow fibers for heavy metal removal

Dye‐affinity adsorption has been used increasingly for heavy metal removal. Synthetic hollow fibers have advantages as support matrices in comparison to conventional bead supports because they are not compressible and they eliminate internal diffusion limitations. The goal of this study was to investigate in detail the performance of hollow fibers composed of modified polyamide to which Cibacron Blue F3GA was attached for the removal of heavy metal ions. The Cibacron Blue F3GA loading was 1.2 mmol/g. The internal matrix was characterized by scanning electron microscopy. No significant changes in the hollow fiber cross‐section or outer layer morphology were observed after dye modification. The effect of the initial concentration of heavy metal ions and medium pH on the adsorption efficiency were studied in a batch reactor. The adsorption capacity of the hollow fibers for the selected metal ions [i.e., Cu(II), Zn(II) and Ni(II)] were investigated in aqueous media with different amounts of these ions (10–400 ppm) and at different pH values (3.0–7.0). The maximum adsorptions of metal ions onto the Cibacron Blue F3GA‐attached hollow fibers were 246.2 mg/g for Cu(II), 133.6 mg/g for Zn(II), and 332.7 mg/g for Ni(II). Furthermore, a Langmuir expression was calculated to extend the adsorption equilibrium. Nitric acid (0.1M) was chosen as the desorption solution. High desorption ratios (up to 97%) were observed in all cases. Consecutive adsorption/desorption operations showed the feasibility of repeated use of this novel sorbent system. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 83: 3089–3098, 2002; DOI 10.1002/app.2338     

Preconcentration of phosphate ion onto ion-imprinted polymer

In this study, selective separation and preconcentration of phosphate ions on the phosphate-imprinted chitosan-succinate beads have investigated. Chitosan-succinate, phosphate, epichlorohydrin were used as the complexing monomer, template and crosslinking agent, respectively. In the first step, chitosan was modified with succinic anhydrides and complex formation occurred between carboxylic acid functional groups and iron(III) ions. Secondly, Fe(III)-chitosan-succinate particles were reacted with phosphate ions. Afterwards, particles were crosslinked with epichlorohydrin and the template (phosphate ions) was removed using 1M KOH solution. Selective cavity for the phosphate ion was obtained in the phosphate-imprinted metal-chelate polymer. These phosphate-imprinted metal-chelate polymer was used in the adsorption-desorption process. The adsorption process was fast and equilibrium was reached around 30 min. The adsorption behaviour of this system was described approximately by the Langmuir equation. Percent extraction, distribution ratio and selectivity coefficients of phosphate and other ions using non-imprinted and phosphate-imprinted polymer were also determined and comparison of these data was reported.