A new liquid-phase enzyme immunoassay for rabbit IgM antibodies and its application for comparing the specificity of IgM and IgG antibodies contained in the same antiserum.
A new liquid-phase enzyme immunoassay (EIA) has been developed to compare the specificities of IgM and IgG antibodies when they are both present in the same serum sample and directed towards a simple hapten. The hapten viomycin (VM) was used as the model antigen and antibodies to VM were raised in rabbits. In the immunoassay VM, labelled with the enzyme galactosidase as marker (VM-GAL), was mixed with rabbit anti-VM serum. First IgM anti-VM antibodies bound to VM-GAL were precipitated with a guinea pig anti-rabbit IgM serum and galactosidase activity was measured in the precipitate. Then IgG anti-VM antibodies bound to VM-GAL were precipitated from the supernatant with a goat anti-rabbit IgG serum and enzyme activity was measured in this precipitate. The guinea pig anti-rabbit IgM and goat anti-rabbit IgG were specific for IgM and IgG respectively and did not appear to cross-react. Nine analogues of VM were used as inhibitors in this immunoassay to compare the specificities of IgM and IgG antibodies for determinants on VM. The results suggest that recognition of the fine structure of VM by IgM is less strict than recognition by IgG.
Tanimori
H
H
Faculty of Pharmaceutical Sciences, Nagasaki University, Japan.
Kitagawa
T
T
D000818:Animals; D000918:Antibody Specificity; D001667:Binding, Competitive; D003429:Cross Reactions; D005260:Female; D006168:Guinea Pigs; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis* / Q000276:immunology; D007075:Immunoglobulin M / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D014756:Viomycin / Q000032:analysis* / Q000276:immunology
D007074:Immunoglobulin G; D007075:Immunoglobulin M; D014756:Viomycin