Characterization of model antigens composed of biotinylated haptens bound to avidin.
Model antigens of defined structure and limited valency have been prepared by binding 2,4-dinitrophenyl (DNP)-biotin haptens of different lengths to the biotin binding sites of avidin. These complexes have been characterized structurally by spectroscopic methods and functionally by testing their ability to crosslink cell-bound anti-DNP immunoglobulin E-receptor complexes to stimulate degranulation of rat basophilic leukemia cells. One of these haptens, 1-DNP-amino-12-biotinamido dodecane, is shown to have only two tight binding sites per avidin molecule, and the resulting bivalent (DNP-biotin)-avidin antigen is found to stimulate substantial degranulation. Another DNP-biotin hapten that is approximately 10 A longer has four tight binding sites per avidin and, when bound to avidin, has greater activity similar to a highly DNP-conjugated multivalent antigen. These and other (DNP-biotin)-avidin complexes described here represent a new group of chemically well-defined antigens that are useful for studying the binding to and functional crosslinking of cell-bound immunoglobulins in immunological responses.
Kane
P
P
Department of Chemistry, Cornell University, Ithaca, NY 14853-1301.
Holowka
D
D
Baird
B
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000941:Antigens / Q000276:immunology*; D001360:Avidin / Q000276:immunology*; D001491:Basophils / Q000378:metabolism; D001710:Biotin / Q000276:immunology*; D004140:Dinitrophenols / Q000276:immunology; D006241:Haptens / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology; D015471:Leukemia, Basophilic, Acute / Q000473:pathology; D051381:Rats; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000941:Antigens; D004140:Dinitrophenols; D006241:Haptens; D001360:Avidin; D007073:Immunoglobulin E; D001710:Biotin