Mehmet Odabaşı

Chitosan microspheres and sponges: Preparation and characterization

In this study, chitosan microspheres and sponges were prepared and characterized for diverse biomedical applications successfully. The chitosan microspheres were obtained with a “suspension crosslinking technique” in the size range of 30–700 μm. The stirring rate of the suspension medium and the chitosan/acetic acid ratio, emulsifier, and crosslinker, that is, the glutaraldehyde concentration in the suspension medium, were evaluated as the effective parameters on the size/size distributions of the microspheres. The microsphere size/size distributions were increased with the decreasing of all effective parameters except the chitosan/acetic acid ratio. In the second part of the study, chitosan sponges were prepared with a solvent-evaporation technique and sponges were cross-linked either during the formation or after the formation of sponges by using a cross-linker, that is, glutaraldehyde. When the sponges were crosslinked during the formation, fibrillar structures were obtained, while the leaflet structures were obtained in the case of crosslinking after the formation of sponges. In the last part of the study, the swelling behavior of both the chitosan microspheres and sponges were evaluated using different amounts of the crosslinker. The swelling ratio was increased in both types of structures, that is, microspheres and sponges, by decreasing the amount of the crosslinker.

Polyhydroxyethylmethacrylate-based magnetic DNA-affinity beads for anti-DNA antibody removal from systemic lupus erythematosus patient plasma

The aim of this study is to prepare magnetic poly(2-hydroxyethylmethacrylate) (mPHEMA) beads and to investigate their utility for the removal of anti-DNA antibodies from systemic lupus erythematosus (SLE) patient plasma. mPHEMA beads, in the size range of 80-120 microm, were produced by a modified suspension technique. Then, DNA was coupled onto mPHEMA beads by carbodiimide activation. The amount of ligand coupled was changed by changing the initial concentrations of carbodiimide and DNA. Human immunoglobulin G (HIgG) and anti-DNA antibody adsorption from aqueous solutions and human plasma were examined in a batch system. mPHEMA beads were characterized by swelling tests, electron spin resonance (ESR) and scanning electron microscopy. Important results obtained in this study are as follows: the swelling ratio of mPHEMA beads was 34%. The presence of magnetite particles in the polymeric structure was confirmed by ESR. The mPHEMA beads have a spherical shape and porous structure. Maximum DNA coupling of carbodiimide activated mPHEMA beads was 4.4 mg/g. Maximum HIgG adsorption from an aqueous solution was 47.5 mg/g. Anti-DNA antibody adsorption from SLE plasma was observed as 87.6 mg/g. Non-specific HIgG adsorption was 0.1 mg/g. More than 90% of the adsorbed HIgG molecules and anti-DNA antibodies were desorbed succesfully by using NaSCN solution. It was possible to reuse these DNA-affinity beads without significant losses in the antibody adsorption capacities.

Human serum albumin chromatography by Cibacron Blue F3GA-derived microporous polyamide hollow-fiber affinity membranes

An affinity dye ligand, Cibacron Blue F3GA was covalently attached onto commercially available microporous polyamide hollow-fibre membranes for human serum albumin (HSA) adsorption from both aqueous solutions and human plasma. Different amounts of Cibacron Blue F3GA were incorporated on the polyamide hollow-fibres by changing the dye attachment conditions, i.e. initial dye concentration, addition of sodium carbonate and sodium chloride. The maximum amount of Cibacron Blue F3GA attachment was obtained at 42.5 micromol g(-1) when the hollow-fibres were treated with 3 M HCI for 30 min before performing the dye attachment. HSA adsorption onto unmodified and Cibacron Blue F3GA-derived polyamide hollow-fibre membranes was investigated batchwise. The non-specific adsorption of HSA was very low (6.0 mg g(-1) hollow-fibre). Cibacron Blue F3GA attachment onto the hollow-fibres significantly increased the HSA adsorption (147 mg g(-1) hollow-fibre). The maximum HSA adsorption was observed at pH 5.0. Higher HSA adsorption was observed from human plasma (230 mg HSA g(-1) hollow-fibre). Desorption of HSA from Cibacron Blue F3GA derived hollow-fibres was obtained using 0. 1 M Tris-HCl buffer containing 0.5 M NaSCN or 1.0 M NaCl. High desorption ratios (up to 98% of the adsorbed HSA) were observed. It was possible to reuse Cibacron Blue F3GA derived polyamide hollow-fibre without significant decreases in the adsorption capacities.

Investigation of lysozyme adsorption performance of Cu2+-attached PHEMA beads embedded cryogel membranes

For the purification of large molecules, cryogels are an alternative stationary phase to particle based media. But, due to the drawbacks of cryogels (i.e., low surface area) and particle sorbents (i.e., pressure drop in fixed beds), in recent years, the use of hybrid cryogels has greatly increased. In this study, a novel hybrid cryogel column was synthesized for the purification of lysozyme from aqueous solutions and hen egg white (HEW). Firstly, poly(2-hydroxyethyl methacrylate) beads (2μm size) were prepared, and after iminodiacetic acid (IDA) immobilization, Cu(2+) ions were attached via the IDA chelating groups. These arranged Cu(2+)-attached PHEMA beads (Cu(2+)-ABs) were located into PHEMA based cryogel in order to prepare Cu(2+)-ABs embedded supermacroporous hybrid cryogel (Cu(2+)-ABsEHC) column. The specific surface area of the hybrid cryogel was found as 95m(2)/g by using BET. The amount of IDA on beads was found as 875μmol IDA/g. It was approached to the optimum adsorption levels at initial lysozyme concentration of 4mg/mL and pH8.0 as 874.9mg/g beads. The purity of lysozyme adsorbed from HEW was studied by SDS-PAGE with a purity of 86.4%. It is demonstrated that this column is a potential separation medium for purification of lysozyme from HEW.

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 μm were produced by suspension polymerization of 2-hydroxyethyl methacrylate (HEMA) and MAC carried out in an aqueous dispersion medium. Then, Fe3+ 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. Fe3+ -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 Fe3+ -chelated poly(HEMA-MAC) beads (Fe3+ 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.

Novel three-dimensional cellulose produced from trunk of Astragalus gummifer (Fabaceae) tested for protein adsorption performance.

This is the first study to produce three-dimensional (3D) cellulose from any plant up to now. This 3D cellulose was produced from Astragalus gummifer(Fabaceae) trunk by using a modified method in which original the shape of cellulose was kept as natural. This novel 3D cellulose was characterized by SEM, TGA, FT-IR, XRD and elemental analysis to evidence the purity and to compare it with commercially available cellulose from cotton. Results from these characterizations were found convincing because almost the same physicochemical properties were observed for both newly obtained 3D cellulose and commercial one. Both fibers and pores on the surface of 3D cellulose were observed. Thanks to its diversified surface morphology, this novel 3D cellulose was tested for its protein adsorption performance and the results were compared with commercial cellulose as follows: maximum adsorption capacity at pH 8.0 was recorded as 59.2 mg/g for 3D cellulose while 29.6 mg/g for commercial cellulose. According to this result, it is clear to say that this sorbent has high affinity for lysozyme. Also this 3D cellulose could be useful for the other areas of separation science.

Magnetic dye-affinity beads for human serum albumin purification.

Cibacron Blue F3GA was covalently attached onto magnetic poly(vinyl alcohol) (mPVAL) beads (100–150 μm in diameter) for human serum albumin (HSA) adsorption from human plasma. Despite low nonspecific adsorption of HSA on mPVAL beads, Cibacron Blue F3GA attachment significantly increased the HSA adsorption. The maximum HSA adsorption was observed at pH 5.0. Higher HSA adsorption was observed from human plasma. Desorption of HSA from mPVAL beads was achieved by medium containing 1.0 M KSCN at pH 8.0. To test the efficiency of albumin adsorption from human serum, before and after albumin adsorption was demonstrated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analyses. HSA molecules could be reversibly adsorbed and desorbed 10 times with the magnetic beads without noticeable loss in their HSA adsorption capacity.

Microsphere-embedded cryogel for selective and efficient depletion of immunoglobulin G from human serum

Abstract In this study a novel composite cryogel column was synthesized for affinity depletion of immunoglobulin G (IgG) from human serum. The microsphere-embedding technique combines the large surface area of microspheres with excellent properties of cryogels. The poly(hydroxyethyl methacrylate- N-methacryloyl-L-histidine methyl ester) microsphere (2 μm size)- embedded composite cryogel (MECC) column was synthesized and characterized by various methods. The specific surface area of the composite cryogel was found as 92 m2/g by using BET measurement. Maximum HIgG adsorption on MECC column was found as 50.5 mg/g. MECC column can be reused many times with no apparent decrease in HIgG adsorption capacity.

Hepatotoxic microcystin removal using pumice embedded monolithic composite cryogel as an alternative water treatment method

Microcystins are the most commonly encountered water-borne cyanotoxins which present short- and long-term risks to human health. Guidelines at international and national level, and legislation in some countries, have been introduced for the effective health risk management of these potent hepatotoxic, tumour-promoters. The stable cyclic structure of microcystins and their common production by cyanobacteria in waterbodies at times of high total dissolved organic carbon content presents challenges to drinking water treatment facilities, with conventional, advanced and novel strategies under evaluation. Here, we have studied the removal of microcystins using three different forms of pumice particles (PPs), which are embedded into macroporous cryogel columns. Macroporous composite cryogel columns (MCCs) are a new generation of separation media designed to face this challenging task. Three different MCCs were prepared by adding plain PPs, Cu(2+)-attached PPs and Fe(3+)-attached PPs to reaction media before the cryogelation step. Column studies showed that MCCs could be successfully used as an alternative water treatment method for successful microcystin removal.

Poly(Styrene‐Hydroxyethyl Methacrylate) Monodisperse Microspheres as Specific Sorbent in Dye Affinity Adsorption of Albumin

Abstract In this study, human serum albumin (HSA) adsorption properties of reactive green HE‐4BD‐attached monodisperse poly(styrene‐2‐hydroxyethyl methacrylate) [poly(St‐HEMA)] microspheres were investigated. Poly(St‐HEMA) microspheres with an uniform size of 4.0 µm in diameter were produced by the dispersion copolymerization of St and HEMA in an ethanol–water medium. Reactive green HE‐4BD was covalently attached onto the poly(St‐HEMA) microspheres via a nucleophilic substitution reaction between the hydroxyl groups of HEMA and triazinyl chloro groups of dye with the equilibrium coupling capacity of 45.8 µmol dye (g polymer)−1. The poly(St‐HEMA) monodisperse microspheres were characterized by scanning electron microscopy. The effect of concentration of HSA, medium pH, and ionic strength on the adsorption efficiency of dye‐attached microspheres were studied in a batch system. The nonspecific adsorption of HSA on the poly(St‐HEMA) microspheres was 2.1 mg g−1. Reactive green HE‐4BD attachment significantly increased the HSA adsorption up to 221 mg g−1. The langmuir adsorption model was found to be applicable in interpreting HSA adsorption by reactive green HE‐4BD attached microspheres. Significant amount of the adsorbed HSA (up to 92%) was eluted in 1 hr in the elution medium containing 1.0‐M NaCl. To determine the effects of adsorption conditions on possible conformational changes of HSA structure, fluorescence spectrophotometry was employed. We resulted that poly(St‐HEMA) dye‐affinity microspheres can be applied for HSA adsorption without causing any significant conformational changes. Repeated adsorption/elution processes showed that these dye‐attached monodisperse microspheres are suitable for HSA adsorption.