Immunological-chromatographic analysis.
Tandem immunoaffinity and conventional high-performance liquid chromatography columns, coupled with a switching valve, were used for the analysis of single and multicomponent antigen samples. Immunoaffinity columns were prepared by hydrophobic adsorption or covalent immobilization of poly- or monoclonal antibodies on macroporous poly(styrene-divinylbenzene) packing materials. Capacities of these columns were constant through at least 77 cycles. Macromolecular antigens were analyzed at submicrogram levels. Antigens bound to the immunoaffinity column were desorbed and concentrated on a conventional analytical column. Gradient elution on the analytical column separated the desorbed antigen(s) from interfering species and permitted the analysis of all species which bound to the immunoaffinity column. Immunological-chromatographic analysis was useful for purification and discrimination of polypeptides of similar three dimensional structures, such as several lysozyme variants.
Janis
L J
LJ
Department of Chemistry, Purdue University, West Lafayette, IN 47907.
Regnier
F E
FE
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D007120:Immunochemistry; D007202:Indicators and Reagents; D011506:Proteins / Q000302:isolation & purification*
D007202:Indicators and Reagents; D011506:Proteins