Ender Unsal

DNA adsorption onto polyethylenimine-attached poly( p-chloromethylstyrene) beads

In this study, DNA adsorption properties of polyethylenimine (PEI)-attached poly(p-chloromethylstyrene) (PCMS) beads were investigated. Spherical beads with an average size of 186 microm were obtained by the suspension polymerization of p-chloromethylstyrene conducted in an aqueous dispersion medium. Owing to the reasonably rough character of the bead surface, PCMS beads had a specific surface area of 14.1 m2/g. PEI chains could be covalently attached onto the PCMS beads with equilibrium binding capacities up to 208 mg PEI/g beads, via a direct chemical reaction between the amine and chloromethyl groups. After PEI adsorption with 10% (w/w) initial PEI concentration, free amino content of PEI-attached PCMS beads was determined as 0.91 mequiv./g. PEI-attached PCMS beads were utilized as sorbents in DNA adsorption experiments conducted at +4 degrees C in a phosphate buffer medium of pH 7.4. DNA immobilization capacities up to 290 mg DNA/g beads could be achieved with the tried sorbents. This value was approximately 50-times higher relative to the adsorption capacities of previously examined sorbents.

pH-sensitive uniform gel beads for DNA adsorption

Uniform gel beads 3 mm in diameter were obtained by the suspension polymerization of an amine functionalized monomer, N-3-(dimethyl amino)propylmethacrylamide (DMAPM). The polymerization of DMAPM in the form of uniform droplets could be achieved at room temperature in an aqueous dispersion medium by using Ca–alginate gel as the polymerization mold. In this preparation, potassium persulfate/tetramethyl ethylenediamine and sodium alginate/calcium chloride were used as the redox initiator and the stabilizer systems, respectively. The crosslinked DMAPM gel beads exhibited pH-sensitive, reversible swelling–deswelling behavior. The uniform gel beads were also obtained by the copolymerization of DMAPM and acrylamide (AA) in the same polymerization system. Although copolymer gel beads with higher pH sensitivities were obtained with increasing feed concentration of DMAPM, the total monomer conversion decreased. Crosslinked DMAPM and DMAPM–AA copolymer gel beads were utilized as sorbents for DNA adsorption. The gel beads produced with higher DMAPM feed concentration exhibited higher equilibrium DNA adsorption capacity. The DNA equilibrium adsorption capacities up to 50 mg DNA/g dry gel could be achieved with the crosslinked DMAPM gel beads. This value was reasonably higher relative to the previously reported adsorption capacities of known sorbents.

Hydroxyl Functionalized Uniform‐Porous Beads, Synthesis and Chromatographic Use

Uniform‐porous poly(dihydroxypropyl methacrylate‐co‐ethylene dimethacrylate), poly(DHPM‐co‐EDM) particles were synthesized as an alternative packing material for reversed phase chromatography. In the synthesis, poly(glycidyl methacrylate‐ethylene dimethacrylate), poly(GMA‐co‐EDM) particles were obtained by a multi‐stage swelling and polymerization protocol, the so called “modified seeded polymerization”. For this purpose, 2.4 µm polystyrene seed particles were first swollen by dibutyl phthalate (DBP) and then by a monomer mixture including glycidyl methacrylate and ethylene dimethacrylate. The repolymerization of monomer phase in the swollen seed particles provided porous uniform particles approximately 7 µm in size. Poly(DHPM‐co‐EDM) particles were obtained by the acid hydrolysis of the particles synthesized with different GMA feed concentrations. These particles were used as column‐packing material in the reversed phase separation of alkylbenzenes. The retention factor‐acetonitrile concentration diagrams clearly showed that the polarity of packing material could be controlled by changing the GMA feed concentration in the “modified seeded polymerization”. The packing materials with more hydrophobic character (i.e., poly(EDM) and poly(DHPM‐co‐EDM) particles produced with the GMA feed concentrations up to 20%) exhibited better chromatographic performance in the reversed phase mode.

Uniform Particles for the Reversed-Phase Separation of Proteins with High-Resolution and High-Column Efficiency

Abstract A low‐sized, uniform and polymer‐based high‐performance liquid chromatography (HPLC) packing material capable of making reversed‐phase separation of proteins with high resolution and with high column efficiency was developed. By a multistage‐swelling and polymerization protocol, 5 µm‐uniform‐porous poly(styrene‐co‐divinylbenzene) particles with relatively larger pores particularly suitable for protein separation were synthesized by starting from a low‐sized seed latex with high average molecular weight and by using a diluent phase comprised of dibutylphthalate and toluene. By the use of synthesized beads as packing material in HPLC, high‐resolution liquid chromatograms were obtained in the gradient separation of selected proteins (i.e., ribonuclease‐A, lysozyme, cytochrome C, and albumin). In the chromatographic runs, the flow rate of the mobile phase was increased fourfold by preserving the resolution power of the column material under gradient conditions. The theoretical plate numbers (TPN) up to 12.500 plates/m were observed by using cytochrome C as the analyte. TPN values determined by the proteins were significantly higher relative to the similar uniform packing materials larger in size (i.e., 7.5–10 µm) obtained by different polymerization methods.