An improved dot immunobinding assay for screening hybridoma supernatants. Non-purified antigen immobilized on nitrocellulose paper discs.
This report describes a modified dot immunobinding assay (DIA) in microplates using a crude mixture of non-purified antigen. Nitrocellulose filter paper discs exposed to the antigen mixture were inserted into the wells and kept in place by a specially constructed device. To test the efficiency of the modification a set of monoclonal antibodies from a mouse immunized with 58 kDa trpE-Bmyc fusion protein were screened. The advantage of this modified method over conventional ELISA is that it permits the use of non-purified antigen for screening large numbers of monoclonal antibodies.
Steinitz
M
M
Department of Tumor Biology, Karolinska Institute, Stockholm, Sweden.
Rosén
A
A
Klein
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D003101:Collodion; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology*; D007118:Immunoassay; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D003101:Collodion