Improvements to the enzyme-developed radial immunodiffusion technique

An enzyme-developed radial immunodiffusion technique, previously known as the diffusion-in-gel enzyme-linked immunosorbent assay (DIG-ELISA), has been improved in two ways: (a) antibody-containing spots have been made larger and more distinct by revealing them with a mixture of hydrogen peroxide, 3,3′-diaminobenzidine and nickel, and further intensification of the ensuing spots with silver; (b) the reliability of the method has been enhanced by chemically coupling the antigen to a layer of a polyamino acid (poly(lysine, phenylalanine)) adsorbed to the bottom of the polystyrene petri dish. The usefulness of the improved technique is illustrated by reference to the measurement of serum concentrations of IgM and IgG, and in the assessment of antibody levels against a particulate antigen (erythrocytes).