Biomedical subjects

Adil Denizli

Publications and source records attributed to Adil Denizli.

31 records · Page 2Linked to original sources

PubMed2005

Immobilized metal affinity beads for ferritin adsorption.

A new metal-chelate adsorbent utilizing N-methacryloyl-(L)-cysteine methyl ester (MAC) was prepared as a metal-chelating ligand. MAC was synthesized by using methacryloyl chloride and L-cysteine methyl ester dihydrochloride. Spherical beads with an average diameter of 150-200 microm were produced by suspension polymerization of 2-hydroxyethyl methacrylate (HEMA) and MAC carried out in an aqueous dispersion medium. Then, Fe(3+) ions were chelated directly on the beads. Properties such as specific surface area, specific pore volume and ligand occupation were determined. The specific surface area of the beads was found to be 18.9 m2/g. The total pore volume was 2.8 ml/g and represented a porosity over 52%. The average pore size of the poly(HEMA-MAC) beads was 620 nm. Fe(3+)-chelated beads were used in the adsorption of ferritin from aqueous solutions. Ferritin adsorption increased with increasing ferritin concentration. The maximum ferritin adsorption capacity of the Fe(3+)-chelated poly(HEMA-MAC) beads (Fe(3+) loading 0.81 mmol/g) was found to be 3.7 mg/g at pH 4.0 in acetate buffer. The non-specific ferritin adsorption on the poly(HEMA-MAC) beads were 0.4 mg/g. Adsorption behavior of ferritin could be modelled using both the Langmuir and Freundlich isotherms. Adsorption capacity decreased with increasing ionic strength of the binding buffer. Ferritin molecules could be adsorbed and desorbed five times with these adsorbents without noticeable loss in their ferritin adsorption capacity.

Cysteine↗

PubMed2004

Molecular recognition based cadmium removal from human plasma.

Molecularly imprinted polymers (MIPs) are easy to prepare, stable, inexpensive and capable of molecular recognition. MIPs can be considered as affinity separation media. Cadmium is a carcinogenic and mutagenic element. There is no specific treatment available for acute or chronic metal poisoning. Besides supportive therapy and hemodialysis, metal poisoning is often treated with commercially available chelating agents including EDTA and dimercaprol. However, there is histopathological evidence for increased toxicity in animals when these agents are utilized. The aim of this study is to prepare ion-imprinted polymers, which can be used for the selective removal of Cd2+ ions from Cd2+-overdosed human plasma. N-Methacryloly-(L)-cysteinemethylester (MAC) was chosen as the complexing monomer. In the first step, Cd2+ was complexed with MAC and the Cd2+ -imprinted p(HEMA-MAC) beads were synthesized by suspension polymerization. After that, the template (i.e., Cd2+ ions) were removed using 0.1 M thiourea solution. The specific surface area of the Cd2+ -imprinted poly(HEMA-MAC) beads was found to be 19.4 m2/g with a size range of 63-140 microm in diameter and the swelling ratio was 78%. According to the elemental analysis results, the beads contained 42.1 micromol MAC/g polymer. The maximum adsorption capacity was 32.5 micromol Cd2+/g beads. The relative selectivity coefficients of imprinted beads for Cd2+/Pb2+ and Cd2+/Zn2+ were 7.8 and 1683 times greater than non-imprinted matrix, respectively. The Cd2+-imprinted poly(HEMA-MAC) beads could be used many times without decreasing their adsorption capacities significantly.

Adsorption↗

PubMed2004

Performance of dye-affinity beads for aluminium removal in magnetically stabilized fluidized bed.

BACKGROUND: Aluminum has recently been recognized as a causative agent in dialysis encephalopathy, osteodystrophy, and microcytic anemia occurring in patients with chronic renal failure who undergo long-term hemodialysis. Only a small amount of Al(III) in dialysis solutions may give rise to these disorders. METHODS: Magnetic poly(2-hydroxyethyl methacrylate) (mPHEMA) beads in the size range of 80-120 microm were produced by free radical co-polymerization of HEMA and ethylene dimethacrylate (EDMA) in the presence of magnetite particles (Fe3O4). Then, metal complexing ligand alizarin yellow was covalently attached onto mPHEMA beads. Alizarin yellow loading was 208 micromol/g. These beads were used for the removal of Al(III) ions from tap and dialysis water in a magnetically stabilized fluidized bed. RESULTS: Al(III) adsorption capacity of the beads decreased with an increase in the flow-rate. The maximum Al(III) adsorption was observed at pH 5.0. Comparison of batch and magnetically stabilized fluidized bed (MSFB) maximum capacities determined using Langmuir isotherms showed that dynamic capacity (17.5 mg/g) was somewhat higher than the batch capacity (11.8 mg/g). The dissociation constants for Al(III) were determined using the Langmuir isotherm equation to be 27.3 mM (MSFB) and 6.7 mM (batch system), indicating medium affinity, which was typical for pseudospecific affinity ligands. Al(III) ions could be repeatedly adsorbed and desorbed with these beads without noticeable loss in their Al(III) adsorption capacity. CONCLUSIONS: Adsorption of Al(III) demonstrate the affinity of magnetic dye-affinity beads. The MSFB experiments allowed us to conclude that this inexpensive sorbent system may be an important alternative to the existing adsorbents in the removal of aluminium.

Journal Article↗

PubMed2004

Concanavalin a immobilized affinity adsorbents for reversible use in yeast invertase adsorption.

Concanavalin A (Con A) immobilized poly(2-hydroxyethyl methacrylate) (PHEMA) beads were investigated for specific adsorption of yeast invertase from aqueous solutions. PHEMA beads were prepared by a suspension polymerization technique with an average size of 150-200 microm, and activated by epichlorohydrin. Con A was then immobilized by covalent binding onto these beads. The maximum Con A immobilization was found to be 10 mg/g. The invertase-loading capability of the PHEMA/Con A beads was 107 mg/g. The maximum invertase adsorption capacity on the PHEMA/Con A adsorbents was observed at pH 5.0. The values of the Michaelis constant K(m) of invertase were significantly larger upon adsorption, indicating decreased affinity by the enzyme for its substrate, whereas V(max) was smaller for the adsorbed invertase. Adsorption improved the pH stability of the enzyme as well as its temperature stability. Thermal stability was found to increase with adsorption. The adsorbed enzyme activity was found to be quite stable in repeated experiments. Storage stability of adsorbed invertase.

Adsorption↗

PubMed2004

Dye affinity hollow fibers for albumin purification.

Reactive Green HE 4BD was immobilized on polyamide (PA) hollow fibers for human serum albumin (HSA) adsorption from both aqueous solutions and human plasma. Different amounts of Reactive Green HE 4BD were incorporated on the PA hollow fibers by changing the dye attachment conditions, i.e. the initial dye concentration and the addition of sodium carbonate and sodium chloride. The maximum amount of dye attachment was obtained as 39.4 micromol x g(-1) when the hollow fibers were treated with 3 M HCl for 30 min before performing the dye attachment. HSA adsorption onto unmodified and dye-attached hollow fibers was investigated batchwise. The non-specific adsorption of HSA was low (6.0 mg/g hollow fiber). Dye attachment onto the hollow fibers significantly increased the HSA adsorption (86.7 mg/g). The maximum HSA adsorption was observed at pH 5.0. Higher HSA adsorption was observed from human plasma (198 mg HSA/g). The desorptions were performed by adding 0.1 M Tris/HCl buffer containing 0.5 M NaSCN or 1.0 M NaCl to the HSA solutions in which adsorption equilibria had been reached. The desorption results demonstrated that the adsorption of HSA to the adsorbent was reversible. Chemical structure of Reactive Green HE-4BD.

Adsorption↗

PubMed2004

Poly(ethylene glycol dimethacrylate-n-vinyl imidazole) beads for heavy metal removal.

Poly(ethylene glycol dimethacrylate-n-vinyl imidazole) [poly(EGDMA-VIM)] hydrogel (average diameter 150-200 microm) was prepared by copolymerizing ethylene glycol dimethacrylate (EGDMA) with n-vinyl imidazole (VIM). The copolymer hydrogel bead composition was characterized by elemental analysis and found to contain 5 EGDMA monomer units each VIM monomer unit. Poly(EGDMA-VIM) beads had a specific surface area of 59.8 m2/g. Poly(EGDMA-VIM) beads were characterized by swelling studies and scanning electron microscopy (SEM). These poly(EGDMA-VIM) beads with a swelling ratio of 78% were used for the heavy metal removal studies. Chelation capacity of the beads for the selected metal ions, i.e., Cd(II), Hg(II) and Pb(II) were investigated in aqueous media containing different amounts of these ions (10-750 mg/l) and at different pH values (3.0-7.0). Chelation rate was very fast. The maximum chelation capacities of the poly(EGDMA-VIM) beads were 69.4 mg/g for Cd(II), 114.8 mg/g for Pb(II) and 163.5 mg/g for Hg(II). The affinity order on molar basis was observed as follows: Hg(II) > Cd(II) > Pb(II). Chelation behavior of heavy metal ions could be modelled using both the Langmuir and Freundlich isotherms. pH significantly affected the chelation capacity of VIM incorporated beads. Chelation of heavy metal ions from synthetic wastewater was also studied. The chelation capacities are 45.6 mg/g for Cd(II), 74.2 mg/g for Hg(II) and 92.5 mg/g for Pb(II) at 0.5 mmol/l initial metal concentration. Regeneration of the chelating-beads was easily performed with 0.1 M HNO3. These features make poly(EGDMA-VIM) beads potential candidate adsorbent for heavy metal removal.

Acrylates↗

PubMed2003

Novel metal-chelate affinity adsorbent for purification of immunoglobulin-G from human plasma.

Metal-chelating ligand and/or comonomer 2-methacrylolyamidohistidine (MAH) was synthesized by using methacryloyl chloride and L-histidine methyl ester. MAH was characterized by NMR and FTIR. Spherical beads with an average diameter of 75-125 microm were produced by suspension polymerization of methylmethacrylate (MMA) and MAH carried out in an aqueous dispersion medium. Poly(MMA-MAH) beads had a specific surface area of 37.5 m(2)/g. Poly(MMA-MAH) beads were characterized by water uptake studies, FTIR, SEM and elemental analysis. Elemental analysis of MAH for nitrogen was estimated as 34.7 microM/g of polymer. Then, Cu(2+) ions were chelated on the beads. Cu(2+)-chelated beads with a swelling ratio of 38% were used in the adsorption of human-immunoglobulin G (HIgG) from both aqueous solutions and human plasma. The maximum adsorption capacities of the Cu(2+)-chelated beads were found to be 12.2 mg/g at pH 6.5 in phosphate buffer and 15.7 mg/g at pH 7.0 in MOPS. Higher adsorption value was obtained from human plasma (up to 54.3 mg/g) with a purity of 90.7%. The metal-chelate affinity beads allowed one-step separation of HIgG from human plasma. The adsorption-desorption cycle was repeated 10 times using the same beads without noticeable loss in their HIgG adsorption capacity.

Adsorption↗

PubMed2003

Immunoaffinity beads for selective removal of cholesterol from human plasma.

Anti-low density lipoprotein antibody (anti-LDL antibody) attached poly(2-hydroxyethyl methacrylate-methacryloylamidophenylalanine) (poly(HEMA-MAPA)) beads were prepared for selective removal of cholesterol from hypercholesterolemic human plasma. Poly(HEMA-MAPA) beads were produced by a modified suspension polymerization and then characterized by swelling tests and SEM. Blood-compatibility tests were also investigated. The water swelling ratio of the poly(HEMA-MAPA) beads increased significantly (68%) compared with pHEMA (55%). All the clotting times increased when compared with poly(HEMA) beads. Loss of platelets and leukocytes was very low. The maximum anti-LDL antibody attachment was achieved at pH 7.0. Attachment of anti-LDL antibody was 29.6 mg/g. There was a very low non-specific cholesterol binding onto the poly(HEMA-MAPA) beads, about 0.74 mg/g. Anti-LDL antibody attached beads adsorbed in the range of 13.3-16.0 mg cholesterol/g from hypercholesterolemic human plasma. Up to 92% of the adsorbed LDL was desorbed. The binding-elution cycle was repeated 10 times using the same beads. There was no significant loss of binding capacity.

Adsorption↗

PubMed2003

N-methacryloly-(L)-histidinemethylester carrying a pseudospecific affinity sorbent for immunoglobulin-G isolation from human plasma in a column system.

N-methacryloly-(L)-histidinemethylester (MAH) as a pseudospecific ligand was synthesized by using methacryloyl chloride and histidine. Spherical beads with an average size of 63-75 microm were obtained by suspension polymerization of ethylene glycol dimethacrylate (EGDMA), 2-hydroxyethyl methacrylate (HEMA) and MAH conducted in an aqueous dispersion medium. The specific surface area of the beads was found to be 18.3 m2/g. Poly(EGDMA-HEMA-MAH) beads were used in the separation of immunoglobulin-G (HIgG) from aqueous solutions and/or human plasma in a packed-bed column system. HIgG adsorption capacity of the beads decreased with an increase in the flow-rate of plasma. The maximum HIgG adsorption on the poly(EGDMA-HEMA-MAH) sorbents was observed at pH 7.4. HIgG adsorption onto the poly(EGDMA-HEMA) sorbents was negligible. Higher adsorption values (up to 135 mg/g) were obtained when the poly(EGDMA-HEMA-MAH) sorbents were used from aqueous solutions. HIgG adsorption increased with decreasing temperature and the maximum adsorption achieved at 4 degrees C. MAH incorporation significantly affected HIgG adsorption capacity (135 mg/g). Higher amounts of HIgG were adsorbed from human plasma (up to 165 mg/g). Adsorption capacities of other blood proteins were obtained as 8.7 mg/g for fibrinogen and 14.6 mg/g for albumin. The total protein adsorption was determined as 191 mg/g. The pseudospecific affinity beads allowed one-step separation of HIgG from human plasma. HIgG molecules could be repeatedly adsorbed and desorbed with these sorbents without noticeable loss in their HIgG adsorption capacity.

Adsorption↗

PubMed2002

Preparation of immuno-affinity membranes for cholesterol removal from human plasma.

Anti-low density lipoprotein antibody (anti-LDL) immobilized polyhydroxyethylmethacrylate (pHEMA) based membrane was prepared for selective removal of cholesterol from hypercholesterolemic human plasma. In order to further increase blood-compatibility, a newly synthesized comonomer, methacryloylamidophenylalanine (MAPA) was included in the membrane formulation. p(HEMA-MAPA) membranes were produced by a photopolymerization and then characterized by swelling tests, SEM and contact angle studies. Blood-compatibility tests were also investigated. The water swelling ratio of the p(HEMA-MAPA) membrane increases significantly (133.2.9%) compared with pHEMA (58%). p(HEMA-MAPA) membranes have large pores around in the range of 5-10 microm. All the clotting times increased when compared with pHEMA membranes. Loss of platelets and leukocytes was very low. The maximum anti-LDL antibody immobilization was achieved around pH 7.0. Immobilization of anti-LDL antibody was 12.6 mg/ml. There was a very low non-specific cholesterol adsorption onto the plain p(HEMA-MAPA) membranes, about 0.36 mg/ml. Anti-LDL antibody immobilized membranes adsorbed in the range of 4.5-7.2 mg cholesterol/ml from hypercholesterolemic human plasma. Up to 95% of the adsorbed LDL antibody was desorbed. The adsorption-desorption cycle was repeated 10 times using the same membrane. There was no significant loss in the adsorption capacity.

Adsorption↗

PubMed2002

Poly(hydroxyethyl methacrylate-co-methacrylamidoalanine) membranes and their utilization as metal-chelate affinity adsorbents for lysozyme adsorption.

Different adsorbents have been reported in the literature for protein purification. The authors have developed a novel and new approach to obtain high protein adsorption capacity utilizing a 2-methacrylamidoalanine-containing membrane. Amino acid ligand 2-methacrylamidoalanine (MAAL) monomer was synthesized using methacryloyl chloride and alanine. Poly(2-hydroxyethylmethacrylate-co-2-methacrylamidoalanine) [p(HEMA-co-MAAL)] membranes were then prepared by UV-initiated photopolymerization of HEMA and MAAL in the presence of an initiator (azobisisobutyronitrile, AIBN). The synthesized MAAL monomer was characterized by NMR. p(HEMA-co-MAAL) membranes were characterized by swelling studies, porosimeter, SEM, FTIR, and elemental analysis. These membranes have macropores in the size range of 5-10 microm. Cu(II) ions (25.9 mmol/m2) were chelated on these membranes. p(HEMA-co-MAAL) membranes were used to study the adsorption of lysozyme from aqueous media containing different amounts of lysozyme (0.1-3.0 mg/l) and at different pH values (4.0-8.0). The non-specific adsorption of lysozyme on the pHEMA membranes was negligible (0.9 microg/cm2). Incorporation of MAAL increased the lysozyme adsorption significantly up to 2.96 mg/cm2. The lysozyme adsorption capacity of the Cu(II) incorporated membranes (9.98 mg/cm2) was greater than that of the p(HEMA-co-MAAL) membranes. More than 90% of the adsorbed lysozyme was desorbed in 1 h in the desorption medium containing 1.0 M NaCl and 0.025 M EDTA. The metal-chelate affinity membranes are suitable for repeated use for more than ten cycles without a noticeable loss of capacity.

Adsorption↗

PubMedDate not supplied

A new metal chelate affinity adsorbent for cytochrome C.

We have prepared a novel metal-chelate adsorbent utilizing N-methacryloyl-L-histidine methyl ester (MAH) as a metal-chelating ligand. MAH was synthesized by using methacryloyl chloride and l-histidine methyl ester dihydrochloride. Spherical beads with an average diameter of 75-125 microm were produced by suspension polymerization of 2-hydroxyethyl methacrylate (HEMA) and MAH carried out in an aqueous dispersion medium. Then, Cu(2+) ions were chelated directly on the chelating beads. Cu(2+)-chelated beads were used in the adsorption of cytochrome c (cyt c) from aqueous solutions. The maximum cyt c adsorption capacity of the Cu(2+)-chelated beads (658.2 micromol/g Cu(2+) loading) was found to be 31.7 mg/g at pH 10 in phosphate buffer. The nonspecific cyt c adsorption on the naked PHEMA beads was 0.2 mg/g. Cyt c adsorption increased with increasing Cu(2+) loading. Cyt c adsorption capacity was demonstrated for the buffer types with the effects in the order phosphate > HEPES > MOPS > MES > Tris-HCl. Cyt c molecules could be adsorbed and desorbed five times with these adsorbents without noticeable loss in their cyt c adsorption capacity.

Adsorption↗

PubMedDate not supplied

A novel magnetic adsorbent for immunoglobulin-g purification in a magnetically stabilized fluidized bed.

A novel magnetic poly(ethylene glycol dimethacrylate-N-methacryloly-L-histidinemethylester) [m-poly(EGDMA-(MAH)] support was prepared for purification of immunoglobulin G (IgG) in a magnetically stabilized fluidized bed by suspension polymerization. Elemental analysis of the magnetic beads for nitrogen was estimated as 70 micromol MAH/g polymer. Magnetic poly(EGDMA-MAH) beads were used in the separation of immunoglobulin-G (IgG) from aqueous solutions and/or human plasma in a magnetically stabilized fluidized bed system. IgG adsorption capacity of the beads decreased with an increase in the flow rate. The maximum IgG adsorption was observed at pH 6.0 for MES buffer. IgG adsorption onto the m-poly(EGDMA) was negligible. Higher adsorption values (up to 262 mg/g) were obtained in which the m-poly(EGDMA-MAH) sorbents were used from aqueous solutions. Higher amounts of IgG were adsorbed from human plasma (up to 320 mg/g) with a purity of 87%. IgG molecules could be repeatedly adsorbed and desorbed with these sorbents without noticeable loss in their IgG adsorption capacity.

Adsorption↗