Abstract

This chapter explains the basic principles of competitive and immunometric assays, starting with the kinetics of antibody-antigen binding reactions, based on the Law of Mass Action. The determinants of assay sensitivity are examined in detail, for both competitive and immunometric assays, with simulations of assays to show how sensitivity can best be optimized during the process of immunoassay design. Three-dimensional graphs are used to explain the relationships between antibody concentration, equilibrium constant, specific activity, non-specific binding and assay sensitivity. There are sections on the impact of solid-phases on assay kinetics and considerations for the determination of antibody levels in samples.

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