A sandwich enzyme immunoassay of rabbit immunoglobulin G with an enzyme labeling method and a new solid support.
A method was devised for enzyme labeling goat antibody to rabbit immunoglobulin G with beta-D-galactosidase (EC 3.2.1.23) using a heterobifunctional cross-linker, N-(gamma-maleimidobutyryloxy)-succinimide. Labeling of the purified antibody was by a continuous 2-step process and including a chromatographic purification procedure could be completed within one day. The partially purified anti-rabbit IgG was coated on Amino-Dylark cylinders, a new solid support, using glutaraldehyde as the coupling reagent. With enzyme-labeled antibody and the solid-phase anti-rabbit IgG, a sandwich enzyme immunoassay for rabbit IgG was developed with a lower limit of detection at 3.5 pM (0.1 ng/tube). The specificity of the assay was excellent and all 4 types of IgG tested showed 0.0001% or less cross-reactivity with rabbit IgG.
D000818:Animals; D000936:Antigen-Antibody Complex; D006041:Goats / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis*; D007202:Indicators and Reagents; D013045:Species Specificity; D001616:beta-Galactosidase
D000936:Antigen-Antibody Complex; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D001616:beta-Galactosidase