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Clinical Chemistry 19: 699-705, 1973;
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Clinical Chemistry, Vol 19, 699-705, Copyright © 1973 by the American Association for Clinical Chemistry

Principles of Competitive Binding Assays (Saturation Analyses). I. Equilibrium Techniques

Alfred Zettner 1

1 Division of Clinical Pathology, Department of Pathology, School of Medicine, the University of California, San Diego.

The principles of competitive binding assays are reviewed, with emphasis on the differentiation of equilibrium vs. sequential saturation techniques. Three methods of equilibrium analysis are outlined that differ from each other in the relative concentrations of labeled ligand and binder used in the incubation system. The linearization of dose-response curves through the use of the logit plot is discussed. The importance of the knowledge of the effective affinity constant K for optimization of the assays is stressed, and two methods of estimating K are demonstrated with a digoxin-antidigoxin-antiserum system.


Key Words: sequential saturation techniques • logit plot in curve linearization • radioimmunoassay • estimating affinity constant • Scatchard plot

Submitted on March 30, 1973
Accepted on April 12, 1973




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Copyright © 1973 by the American Association for Clinical Chemistry.