Effective avidin coating of erythrocytes by disulfide bond formation. Production of a stable rosetting agent for multiple applications.
A method is described for covalently coupling avidin to the membrane of glutaraldehyde-treated erythrocytes. The method is based on the use of the heterobifunctional coupling reagent N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and appears to be more effective than other protein coupling techniques. The resulting avidin-coated red cells are very stable and can be stored for more than 1 year without aggregation or loss of biotin-binding activity. They can be used in various rosetting assays and were tested for T cell depletion or negative cell selection with five biotinylated monoclonal antibodies.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D001360:Avidin; D001710:Biotin; D002469:Cell Separation / Q000379:methods; D004220:Disulfides; D004910:Erythrocyte Membrane; D007158:Immunologic Techniques; D010047:Ovalbumin / Q000031:analogs & derivatives*; D011725:Pyridines; D012397:Rosette Formation; D012756:Sheep
D000911:Antibodies, Monoclonal; D004220:Disulfides; D011725:Pyridines; D001360:Avidin; D001710:Biotin; D010047:Ovalbumin