Elimination of undesired cross-reactants by using mixtures of antibodies: experimental and theoretical evaluations of hapten radioimmunoassays

Abstract

Abstract This study shows that the specificity of radioimmunoassays can be improved by including a second antibody raised against an undesired cross-reactant. In a radioimmunoassay of prostaglandin E2 (PGE2) involving a monoclonal antibody, the cross-reactivity with 6-keto-prostaglandin E1 (6kPGE1) was decreased from 20% to 2% by including a high concentration of a polyclonal anti-6kPGE1. A similar increase in specificity was obtained in the assay of a larger hapten, luliberin (luteinizing hormone releasing hormone); the cross-reactivity of a luliberin analog was decreased 20-fold. Equations derived from the Law of Mass Action were used for the mathematical analysis and for the computer simulation of changes in assay affinity and specificity according to the quantity and quality of the mixed antibodies. The model gave values that agreed well with experimental data; it promises to be quite useful in designing specific radioimmunoassays.