title: Immunoaffinity chromatography.
abstract: This unit describes the isolation of soluble or membrane-bound protein antigens from cells or homogenized tissue by immunoaffinity chromatography. The technique involves the elution of a single protein from an immunoaffinity column after prior elution of nonspecifically adsorbed proteins. To elute the bound antigen from the immunoaffinity matrix, the antibody-antigen interaction is destabilized by brief exposure to high-pH or low-pH buffer. The use of batch purification of antigens is an alternate procedure and results in shorter column loading times. The detergent octyl beta-D-glucoside can be used instead of Triton X-100 for elution. Because octyl beta-D-glucoside has a high critical micelle concentration (CMC), it can be readily removed by dialysis, as described.
authors.affiliation: Center for Blood Research, Harvard Medical School, Boston, Massachussets.
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Springer
mesh_terms: D000906:Antibodies / Q000737:chemistry; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D001345:Autoradiography / Q000379:methods; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods*; D003902:Detergents / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006863:Hydrogen-Ion Concentration; D047468:Immunoprecipitation; D011506:Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D000906:Antibodies; D000941:Antigens; D003902:Detergents; D011506:Proteins