The choice of methods for immunoglobulin IgG purification: yield and purity of antibody activity.
Six methods for the purification of immunoglobulin G (IgG) from serum were compared, using rabbit antiserum to Bacillus anthracis spores as a model. Antibody activity was monitored by a solid-phase immunoradiometric assay (IRMA). Salt precipitation/ion exchange chromatography and ethanol precipitation both resulted in IgG of high purity but there was considerable inactivation of antibody. Salt precipitation/affinity chromatography gave poor yields of antibody. PEG precipitation and gel filtration of Sephacryl S-300 gave moderate yields and purity of IgG, with little evidence of antibody inactivation. Salt precipitation was marginally more destructive than the last 2 methods, but is recommended for routine use on grounds of its simplicity. Should IgG prepared by salt precipitation prove inadequate for particular applications, gel filtration is recommended since it allows the balance of yield and purity to be altered at will.
D000818:Animals; D001408:Bacillus anthracis / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D004588:Electrophoresis, Cellulose Acetate; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods; D013171:Spores, Bacterial / Q000276:immunology
D007074:Immunoglobulin G