A new process of IgG purification by negative chromatography: adsorption aspects of human serum proteins onto omega-aminodecyl-agarose.
The adsorbent omega-aminodecyl-agarose was evaluated as to its feasibility for the adsorption of human serum and plasma proteins, aiming at the purification of immunoglobulin G (IgG). The contribution of electrostatic and hydrophobic interactions (mixed-mode) and the effects of buffer system on the adsorption of serum proteins were also studied. The adsorption isotherm parameters of human serum albumin (HSA) and IgG were evaluated, pointing to the existence of cooperative effects in the process. A positive (n=2.30+/-0.38) and negative cooperativity (n=0.63+/-0.12) were observed for IgG and HSA binding, respectively. High purity IgG was obtained (based on total protein concentration and nephelometric analysis of HSA, transferrin, and immunoglobulins A, G, and M) with a 75% recovery in Hepes 25 mmol L(-1) pH 6.8 feeding human serum. These results indicate that the use of omega-aminodecyl-agarose is a potential technique for purification of IgG from human serum.
Bresolin
Igor Tadeu Lazzarotto
IT
Department of Biotechnological Processes, School of Chemical Engineering, University of Campinas, UNICAMP, P.O. Box 6066, 13083-970 Campinas, SP, Brazil.
de Souza
Maria Cristiane Martins
MC
Bueno
Sonia Maria Alves
SM
D000327:Adsorption; D001798:Blood Proteins / Q000302:isolation & purification*; D002021:Buffers; D002845:Chromatography / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007700:Kinetics; D008956:Models, Chemical; D009391:Nephelometry and Turbidimetry; D010084:Oxidation-Reduction; D011485:Protein Binding; D012685:Sepharose / Q000031:analogs & derivatives* / Q000737:chemistry*; D044967:Serum; D012709:Serum Albumin / Q000302:isolation & purification; D012996:Solutions; D013696:Temperature
D001798:Blood Proteins; D002021:Buffers; D007074:Immunoglobulin G; D012709:Serum Albumin; D012996:Solutions; D012685:Sepharose