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Clinical Chemistry 21: 1769-1773, 1975;
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Clinical Chemistry, Vol 21, 1769-1773, Copyright © 1975 by the American Association for Clinical Chemistry

Scatchard Plot and Heterogeneity in Binding Affinity of Labeled and Unlabeled Ligand

H. J. G. Hollemans 1 and R. M. Bertina 1

1 School of Medicine, Division of Endocrinology, University Hospital Wilhelmina Gasthuis, Eerste Helmersstraat 104, University of Amsterdam, The Netherlands.

In saturation analysis the Scatchard plot is a generally accepted method for calculation of the affinity constant, K, and the molar concentration, q, of the binder. However, in a system where the K's for the labeled and unlabeled ligand are unequal, a nonlinear plot can be obtained from which incorrect values for K and q may be calculated. This paper mathematically explains how the plot may deviate and under which conditions there will be a maximum in the curve. When the binding sites are homogeneous, the coordinates of this maximum can be used to calculate K and q. A general mathematical expression is derived on the basis of which a linear curve can be constructed for calculation of q and K, which is valid even when affinity for the labeled and unlabeled ligand is not identical.

Submitted on April 29, 1975
Accepted on August 21, 1975




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[Abstract] [PDF]


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Copyright © 1975 by the American Association for Clinical Chemistry.