N‐acetyl‐D‐galactosamine‐specific lectin isolation from soyflour with poly(HPMA‐GMA) beads

Soybean lectin was purified from seeds of Glycine max L.Merrill SA88. Poly(hydroxypropyl methacrylate‐glycidyl methacrylate) [poly(HPMA‐GMA)] beads were used as an affinity matrix and N‐acetyl‐D ‐galactosamine (GalNAc) was used as an affinity ligand. Soybean lectin adsorption with GalNAc attached poly(HPMA‐GMA) beads from soybean lectin solution (in phosphate buffered saline) was 5.0 mg/g. Maximum adsorption capacity for soybean lectin from the soy flour extract was 26.0 mg/g. Elution of soybean lectin from adsorbent was accomplished by 0.5M galactose solution. Purity of soybean lectin was determined by SDS‐PAGE. It was observed that soybean lectin could be repeatedly adsorbed and desorbed with GalNAc‐attached poly(HPMA‐GMA) beads. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Dye‐affinity microbeads for removal of phenols and nitrophenols from aquatic systems

Alkali Blue 6B attached poly(2‐hydroxyethyl methacrylate) (PHEMA) microbeads were investigated as dye‐affinity adsorbents for the removal of phenol and nitrophenols (i.e., 2‐nitrophenol, 4‐nitrophenol, and 2,4‐dinitrophenol) from aqueous solutions. PHEMA microbeads were prepared by radical‐suspension polymerization of HEMA in the presence of azobisisobutyronitrile as the initiator. These microbeads with a swelling ratio of 55% and carrying 23.6 μmol Alkali Blue 6B/g polymer were then used in the removal of phenol and nitrophenols from aqueous media. The adsorption was fast in all cases (20‐min equilibrium time). The maximum adsorptions of phenols onto the microbeads carrying Alkali Blue 6B were 145.2 μmol/g for phenol, 87.8 μmol/g for 2,4‐dinitrophenol, 112.6 μmol/g for 4‐nitrophenol, and 104.3 μmol/g for 2‐nitrophenol. The affinity order was phenol > 4‐nitrophenol > 2‐nitrophenol > 2,4‐dinitrophenol. The adsorption of nitrophenols decreased with increasing pH. Desorption of nitrophenols was achieved using a 30% (v/v) methanol solution. The microbeads carrying Alkali Blue 6B are suitable for repeated use for more than five cycles without a noticeable loss of adsorption capacity. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 83: 2411–2418, 2002     

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     

Antibody purification using porous metal–chelated monolithic columns

A novel monolithic material was developed to obtain efficient and cost‐effective purification of IgG from human plasma. The porous monolith was obtained by bulk polymerization in a glass tube of 2‐hydroxyethyl methacrylate (HEMA) and N‐methacryloyl‐(L )‐histidine methyl ester (MAH). The poly(HEMA‐MAH) monolith had a specific surface area of 214.6 m²/g and was characterized by swelling studies, porosity measurement, FTIR, scanning electron microscopy, and elemental analysis. Then the monolith was loaded with Cu²⁺ ions to form the metal chelate. Poly(HEMA‐MAH) monolith with a swelling ratio of 74% and containing 20.9 μmol MAH/g was used in the adsorption/desorption of IgG from aqueous solutions and human plasma. The maximum adsorption of IgG from an aqueous solution in phosphate buffer was 10.8 mg/g at pH 7.0. Higher adsorption was obtained from human plasma (up to 104.2 mg/g), with a purity of 94.1%. It was observed that IgG could be repeatedly adsorbed and desorbed with the poly(HEMA‐MAH) monolith without significant loss of adsorption capacity. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 101: 395–404, 2006     

Metal‐chelating properties of poly(2‐hydroxyethyl methacrylate–methacryloylamidohistidine) membranes

Metal‐chelating membranes have advantages as adsorbents in comparison with conventional beads because they are not compressible and they eliminate internal diffusion limitations. The aim of this study was to explore in detail the performance of poly(2‐hydroxyethyl methacrylate–methacryloylamidohistidine) [poly(HEMA–MAH)] membranes for the removal of three toxic heavy‐metal ions—Cd(II), Pb(II), and Hg(II)—from aquatic systems. The poly(HEMA–MAH) membranes were characterized with scanning electron microscopy and ¹H‐NMR spectroscopy. The adsorption capacity of the poly(HEMA–MAH) membranes for the selected heavy‐metal ions from aqueous media containing different amounts of these ions (30–500 mg/L) and at different pH values (3.0–7.0) was investigated. The adsorption capacity of the membranes increased with time during the first 60 min and then leveled off toward the equilibrium adsorption. The maximum amounts of the heavy‐metal ions adsorbed were 8.2, 31.5, and 23.2 mg/g for Cd(II), Pb(II), and Hg(II), respectively. The competitive adsorption of the metal ions was also studied. When the metal ions competed, the adsorbed amounts were 2.9 mg of Cd(II)/g, 14.8 mg of Pb(II)/g, and 9.4 mg of Hg(II)/g. The poly(HEMA–MAH) membranes could be regenerated via washing with a solution of nitric acid (0.01M). The desorption ratio was as high as 97%. These membranes were suitable for repeated use for more than three adsorption/desorption cycles with negligible loss in the adsorption capacity. The stability constants for the metal‐ion/2‐methacryloylamidohistidine complexes were calculated to be 3.47 × 10⁶, 7.75 × 10⁷, and 2.01 × 10⁷ L/mol for Cd(II), Pb(II), and Hg(II) ions, respectively, with the Ruzic method. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 97: 1213–1219, 2005     

Spectral characterization of lysozyme adsorption on dye-affinity beads

Cibacron Blue F3GA-attached magnetic poly(2-hydroxyethyl methacrylate) [mPHEMA] beads were prepared by suspension polymerization of HEMA in the presence of magnetite (Fe₃O₄) nanopowder. Average diameter size of the mPHEMA beads was 150–200 μm. The characteristic functional groups of Cibacron Blue F3GA-attached mPHEMA beads were analyzed by Fourier transform infrared spectrometer (FTIR) and Raman scattering spectrometer. The lysozyme adsorption and desorption characteristics of Cibacron Blue F3GA-attached mPHEMA beads were also investigated using FTIR and Raman spectroscopic techniques. When the Raman spectrum of lysozyme adsorbed mPHEMA is evaluated characteristic Amide-I band appears at 1657 cm⁻¹. The intensity of this band decreases in the spectrum of lysozyme desorbed mPHEMA sample. When the characteristic bands of lysozyme adsorbed and desorbed mPHEMA samples are compared, the band intensities of desorbed sample are lower than those of lysozyme adsorbed sample except for the band appearing at 656 cm⁻¹ (Tyr vC S). © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008     

Monosize magnetic hydrophobic beads for lysozyme purification under magnetic field

Monosize and magnetic poly(glycidyl methacrylate-N-methacryloyl-(l)-tryptophan) [mPGMATrp] beads (1.6 µm in diameter) were used for hydrophobic affinity capture of lysozyme from chicken egg-white. N-methacryloyl-(l)-tryptophan (MATrp), which gives hydrophobicity to the resulting polymer, was synthesized by reacting methacryloyl chloride and l-tryptophan methyl ester then characterized by Nuclear Magnetic Resonance (NMR). mPGMATrp beads were produced by dispersion polymerization in the presence of magnetite nano-powder. mPGMATrp beads were characterized by means of swelling studies, elemental analysis, Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscope (SEM). Lysozyme adsorption experiments were performed under different experimental conditions (i.e., lysozyme concentration, temperature, and ionic strength) in magnetically stabilized fluidized bed system, (MSFB). Maximum adsorption capacity was 263.9 mg/g. It was observed that mPGMATrp beads can be used without significant loss in lysozyme adsorption capacity after 25 adsorption-elution cycle.     

Albumin adsorption from aqueous solutions and human plasma in a packed-bed column with Cibacron Blue F3GA-Zn(II) attached poly(EGDMA-HEMA) microbeads

Affinity dye-ligand Cibacron Blue F3GA, was covalently coupled with poly(EGDMA-HEMA) microbeads via nucleophilic reaction between the chloride of its triazine ring and the hydroxyl groups of the HEMA under alkaline conditions. The microbeads carrying 16.5 μmol Cibacron Blue F3GA per gram polymer was incorporated with Zn(II) ions. Zn(II) loading was 189.6 μmol/g. Cibacron Blue F3GA-Zn(II) attached affinity sorbent was used for albumin adsorption from aqueous solutions and human plasma in a packed-bed column. BSA adsorption capacity of the microbeads decreased with an increase in the recirculation rate. High adsorption rates were observed at the beginning, then equilibrium was gradually achieved in about 60 min. The BSA concentration in the mobile phase also effected adsorption. BSA adsorption was first increased with BSA concentration, then reached a plateau which was about 128 mg BSA/g. The maximum adsorption was observed at pH 5.0 which is the isoelectric pH of BSA. Higher human serum albumin adsorption was observed from human plasma (215 mg HSA/g). High desorption ratios (over 90% of the adsorbed albumin) were achieved by using 1.0 M NaSCN (pH 8.0) in 30 min.