ovalbumin and ethylene-dimethacrylate

A phenylboronate affinity monolith was prepared and applied to the selective capture of glycoproteins from unfractionated protein mixtures. The monolith was synthesized in a 100 μm i.d capillary by an in situ polymerization procedure using a pre-polymerization mixture consisting of 4-vinylphenylboronic acid (VPBA) as functional monomer, ethylene dimethacrylate (EDMA) as crosslinker, diethylene glycol and ethylene glycol as binary porogenic solvents, and azobisisobutyronitrile (AIBN) as initiator. The prepared monolith was characterized in terms of the morphology, pore property, and recognition property. The selectivity and dynamic binding capacity were evaluated by using standard glycoproteins and nonglycoproteins as model proteins. The chromatographic results demonstrated that the phenylboronate affinity monolith had higher selectivity and binding capacity for glycoprotein than nonglycoprotein. The resulting phenylboronate affinity monolith was used as the sorbent for in-tube solid phase microextraction (in-tube SPME), and the extraction performance of the monolith was assessed by capture of ovalbumin from egg white sample.

Topics: Animals; Boronic Acids; Chickens; Chromatography, Affinity; Chromatography, High Pressure Liquid; Methacrylates; Ovalbumin; Polymers; Solid Phase Microextraction; Vinyl Compounds

A continuous rod of porous poly(glycidyl methacrylate-co-ethylene dimethacrylate) has been prepared by a free radical polymerization within the confines of a 300 x 8 mm I.D. chromatographic column. The epoxide groups of the rod have been modified by a reaction with diethylamine that affords ionizable functionalities required for the ion-exchange chromatographic mode. The properties of this rod column have been characterized and the column has been used successfully for the chromatographic separation of proteins. The column exhibits a dynamic capacity that exceeds 300 mg at a flow velocity of 200 cm/min. An excellent selectivity allows the separation of up to 300 mg of a protein mixture in a single run.

Topics: Animals; Chickens; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Conalbumin; Epoxy Compounds; Methacrylates; Muramidase; Ovalbumin; Proteins; Serum Albumin, Bovine; Trypsin Inhibitors