Choice of immunoglobulin G purification method in assays for antibodies to the thyrotropin receptor.
Activity of autoantibodies to the thyrotropin receptor in the serum of patients with active Graves's disease was compared when the patients' IgG was purified by three different procedures: ammonium sulfate precipitation (I), a modified batch diethylaminoethyl cellulose method (II), and affinity chromatography on Protein A-Sepharose CL-4B (III). IgG extracted by I was significantly less potent in inhibiting binding of 125I-labeled thyroid membranes than that prepared by either II or III, and was significantly less effective than II in stimulating adenyl cyclase activity in thyroid membrane. Thyroglobulin, a serum protein whose concentration is increased in patients with various thyroid diseases, was coprecipitated in amounts sufficient to significantly inhibit binding only when method I was used, but not with either of the other two procedures. Evidently method I is inferior to either of the other two when used for purification of autoantibodies to the thyrotropin receptor. Method II used in this study, being faster and more economical than I and of equivalent efficacy, is a feasible alternative method for clinical use.
D000645:Ammonium Sulfate; D000906:Antibodies / Q000302:isolation & purification*; D001323:Autoantibodies / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D006111:Graves Disease / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000302:isolation & purification*; D018828:Immunoglobulins, Thyroid-Stimulating; D011989:Receptors, Thyrotropin / Q000276:immunology*; D013205:Staphylococcal Protein A; D013961:Thyroid Gland / Q000378:metabolism; D013972:Thyrotropin / Q000037:antagonists & inhibitors
D000906:Antibodies; D001323:Autoantibodies; D007074:Immunoglobulin G; D018828:Immunoglobulins, Thyroid-Stimulating; D011989:Receptors, Thyrotropin; D013205:Staphylococcal Protein A; D013972:Thyrotropin; D000645:Ammonium Sulfate