Factors contributing to intrinsic loading capacity in silica-based packing materials for preparative anion-exchange protein chromatography.
Properties of the matrix and stationary phase which affect the intrinsic loading capacity of silica-based packing materials for preparative anion-exchange chromatography of proteins were investigated. Polyethyleneimine-coated controlled porosity glass beads ranging from 100 to 2000 A in pore diameter were used to evaluate the effects of pore diameter and surface area. Protein binding was found to depend on accessible, rather than total, support surface area. Consequently, wide-pore, high surface area media provide maximum intrinsic loading capacity. Increasing the number of positively charged sites on the stationary phase by increased coating or by quaternization of amines increases hemoglobin-binding capacity.
D000588:Amines; D002852:Chromatography, Ion Exchange / Q000295:instrumentation*; D006454:Hemoglobins / Q000302:isolation & purification*; D006801:Humans; D007202:Indicators and Reagents; D008024:Ligands; D008970:Molecular Weight; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification*; D012822:Silicon Dioxide; D013499:Surface Properties
D000588:Amines; D006454:Hemoglobins; D007202:Indicators and Reagents; D008024:Ligands; D011506:Proteins; D012822:Silicon Dioxide