title: Protein A immobilized affinity cartridge for immunoglobulin purification.
abstract: Recombinant Protein A was immobilized on a cellulose and acrylic composite matrix through Schiff base formation. Various factors that could affect the binding of immunoglobulin by the Protein A molecules immobilized on the solid matrix were studied to achieve optimum affinity purification. The spacer arm length and ligand concentration of Protein A were verified as factors crucial to optimized IgG purification. Liquid-phase environmental conditions such as pH and salt concentration also play important roles in adsorption capacity by affecting the molecular interaction between IgG and the immobilized Protein A. The rate of interaction between Protein A and IgG is rather fast, with minimal differences observed at 10-fold increases in the cartridge loading rate. This paper describes a cellulose/acrylic composite matrix for immobilizing Protein A, at an optimized ligand concentration, installed on a spacer arm of adequate length, to purify immunoglobulins from animal plasma. The fast-flow property of the cartridge made from such a matrix and its simplicity in operation provide effective means for purifying immunoglobulins on a relatively large scale.
authors.affiliation: Life Science Division, Cuno Inc., Meriden, Connecticut 06450.
authors.forename: K C
authors.initials: KC
authors.lastname: Hou
authors.forename: R
authors.initials: R
authors.lastname: Zaniewski
authors.forename: S
authors.initials: S
authors.lastname: Roy
mesh_terms: D000179:Acrylates; D000327:Adsorption; D002482:Cellulose; D002846:Chromatography, Affinity / Q000295:instrumentation*; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000302:isolation & purification; D007136:Immunoglobulins / Q000302:isolation & purification*; D007700:Kinetics; D008024:Ligands; D012545:Schiff Bases; D013205:Staphylococcal Protein A
chemical_list: D000179:Acrylates; D007074:Immunoglobulin G; D007136:Immunoglobulins; D008024:Ligands; D012545:Schiff Bases; D013205:Staphylococcal Protein A; D002482:Cellulose