A biotin-labelled antigen radioimmunoassay (BILA) for antibodies to membrane antigens useful for monoclonal antibody screening.
Viable cells or protein extracts were labelled with N-hydroxysuccinimidobiotin and used as target antigens in a biotin-labelled antigen radioimmunoassay (BILA). The binding of the biotinylated antigens to capture antibodies coated on the bottoms of 96-well plastic plates were measured using 125I-labelled streptavidin as the detection step. The assay circumvents some of the problems associated with solid-phase RIA and permits screening of antibodies to undefined protein antigens present in very small amounts in complex protein solutions.
Jansson
B
B
Department of Tumor Immunology, Wallenberg Laboratory, Lund, Sweden.
Brodin
T
T
Sjögren
H O
HO
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000276:immunology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D001710:Biotin / Q000031:analogs & derivatives*; D002460:Cell Line; D003902:Detergents; D006801:Humans; D051379:Mice; D010047:Ovalbumin / Q000276:immunology; D011863:Radioimmunoassay / Q000379:methods; D051381:Rats; D011920:Rats, Inbred WF; D013388:Succinimides
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D003902:Detergents; D013388:Succinimides; C033788:biotinyl N-hydroxysuccinimide ester; D001710:Biotin; D010047:Ovalbumin