title: Purification of human IgG by negative chromatography on omega-aminohexyl-agarose.
abstract: The omega-aminohexyl diamine immobilized as ligand on CNBr- and bisoxirane-activated agarose gel was evaluated for the purification of human immunoglobulin G (IgG) from serum and plasma by negative affinity chromatography. The effects of matrix activation, buffer system, and feedstream on recovery and purity of IgG were studied. A one-step purification process using Hepes buffer at pH 6.8 allowed a similar recovery (69-76%) of the loaded IgG in the nonretained fractions for both matrices, but the purity was higher for epoxy-activated gel (electrophoretically homogeneous protein with a 6.5-fold purification). The IgG and human serum albumin (HSA) adsorption equilibrium studies showed that the adsorption isotherms of IgG and HSA obeyed the Langmuir-Freundlich and Langmuir models, respectively. The binding capacity of HSA was high (210.4 mg mL(-1) of gel) and a positive cooperativity was observed for IgG binding. These results indicate that immobilizing omega-aminohexyl using bisoxirane as coupling agent is a useful strategy for rapid purification of IgG from human serum and plasma.
authors.affiliation: Department of Biotechnological Processes, School of Chemical Engineering, University of Campinas, UNICAMP, P.O. Box 6066, CEP 13083-970, Campinas, SP, Brazil.
authors.forename: Maria Cristiane Martins
authors.initials: MC
authors.lastname: de Souza
authors.forename: Igor Tadeu Lazzarotto
authors.initials: IT
authors.lastname: Bresolin
authors.forename: Sonia Maria Alves
authors.initials: SM
authors.lastname: Bueno
mesh_terms: D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000378:metabolism; D008024:Ligands; D011485:Protein Binding; D012685:Sepharose / Q000031:analogs & derivatives* / Q000737:chemistry; D012709:Serum Albumin / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D007074:Immunoglobulin G; D008024:Ligands; D012709:Serum Albumin; C024903:aminohexyl-sepharose; D012685:Sepharose