Modified Diffusion in Gel - Enzyme Linked Immunosorbent Assay, (DIG-ELISA) for Quantitation of Specific Antibody

摘要

A method was developed to quantify specific antibody by immobilizing an antigen in an agar film. In our model system, adenovirus types 4 and 7 in 1% agar was allowed to dry to a thin film in 15 x 60 mm plastic tissue culture dishes. This antigen containing film was used to capture specific antibody. Specific antiserum was allowed to diffuse from wells in a gel layered over the antigen containing agar film. After the antibody had diffused, the agar layer was removed and alkaline phosphate conjugated anti rabbit antiserum was layered over the plate. After incubation, this was removed and the plate was layered with a substrate containing agarose. Within 45 minutes, zones of reactivity were visualized with the aid of oblique light. The method proved to be sensitive and end points could be accurately identified with the naked eye. (Author)