ovalbumin and glycidyl-methacrylate

In this study, a novel phosphate-Zr(4+) immobilized metal affinity membrane (IMAM) was prepared based on the surface initiated-atom transfer radical polymerization technique for the selective adsorption of phosphoprotein. The adsorption capacity and selectivity of the phosphate-Zr(4+) IMAM were evaluated by using the mixture of standard phosphoproteins (β-casein, ovalbumin) and nonphosphoproteins (bovine serum albumin and lysozyme) as model samples. The adsorption isotherms and competitive adsorption results demonstrated that the phosphate-Zr(4+) IMAM had higher binding capacity and selectivity for phosphoproteins over nonphosphoproteins. Moreover, the phosphate-Zr(4+) IMAM exhibited good re-usability and re-productivity. Finally, the phosphate-Zr(4+) IMAM was applied to separate phosphoprotein from real samples with high purity. Therefore, the as-prepared phosphate-Zr(4+) IMAM could be a promising affinity material for the efficient enrichment of phosphoprotein from complex bio-samples.

Topics: Adsorption; Animals; Caseins; Cations; Chemical Fractionation; Chickens; Egg Proteins; Egg White; Epoxy Compounds; Membranes, Artificial; Methacrylates; Ovalbumin; Phosphates; Phosphoproteins; Polymerization; Zirconium

Surface grafting of glycidyl methacrylate (GMA) on silica gel and a polyethylene bead was performed by radical polymerization and radiation-induced polymerization, respectively, in order to improve softness. Subsequently, diethylene triamine (DETA), triethylene tetraamine (TETA), and iminodiacetic acid (IDA) were introduced to the grafted GMA for use as affinity columns. The efficiency of the affinity column was investigated by use of bovine serum albumin (BSA) and hemoglobin (Hb) as model proteins. The affinity degree of BSA was higher than Hb for the DETA and TETA column, whereas the affinity degree of Hb was higher than BSA for the IDA column supported by silica gel. The affinity degree of BSA was higher than Hb for the DETA and TTA column supported by polyethylene (PE) beads.

Topics: Amines; Animals; Cattle; Chromatography, Affinity; Epoxy Compounds; Hemoglobins; Hydrogen-Ion Concentration; Imino Acids; Methacrylates; Microspheres; Ovalbumin; Piperazines; Polyamines; Polyethylene; Serum Albumin, Bovine; Silica Gel; Silicon Dioxide; Surface Properties; Time Factors

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