ABSTRACT
High-performance immunoaffinity chromatography (HPIAC) can be used to isolate any biological material or chemical to which an antibody can be made. The membranes were passed through the same HPIAC column under identical conditions and demonstrated similar results, to those found with the lymphocytes, although the second peak developed during this run was smaller. The chapter shows that the adherent cells were recovered by gentle sonication and the membrane-rich fraction obtained in a similar manner. The size of the isolated receptor was the same in both the lymphocyte and macrophage membrane preparations. Membrane proteins isolated by the different HPIAC techniques were analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Isolation of the component 3b receptors showed the presence of a single band of approximately 120 kDa, when compared to the molecular weight markers. Immunoaffinity has been used by several groups as a sample preparation technique prior to conventional high-performance liquid chromatography or in tandem with reversed-phase analysis of the immunoaffinity isolated materials.
