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Methods in the Quantitation of Antigen-Antibody Reactions Measured by Radioimmunoassay: Application to the Gastrin System Revealing Large and Restricted Binding Energies¹

Division of Gastroenterology, Department of Medicine, Nassau County Medical Center, East Meadow, New York 11554; School of Medicine, Health Science Center, State University of New York at Stony Brook, Stony Brook, New York; and Department of Physiology, The Mount Sinai Medical and Graduate Schools of the City University of New York, New York, New York

Abstract

Quantitative methods are presented for defining antigen-antibody interactions in the radioimmunologic assay system when only the bound to free ratio of labeled antigen and the concentration of unlabeled antigen are known. From these methods the equilibrium constant (K), the antibody site concentration (Ab_T), the number of binding sites (n), and the extent of binding energy heterogeneity () are derived for the gastrin-antiporcine gastrin antibody interaction by using human synthetic gastrin for both the labeled and unlabeled antigen. In one method, models of various orders of interaction are applied to the data and suggest a univalent homogeneous gastrin-antibody reaction. In two other methods a form of the Scatchard equation and Sips distribution are applied to the experimental data to obtain an n of one and an close to one, respectively. Values for K are found by all three methods. The interaction was studied at six different antibody dilutions and the equilibrium constant and effective antibody site concentration at the largest dilution were 2.71 x 10¹¹ M^-1 and 1.09 x 10^-12 M, respectively.

Footnotes

¹ This work was supported in part by general research funds from the Meadowbrook Medical Education and Research Foundation and the People of the County of Nassau. Presented in part before the American Society of Biological Chemists at the 57th meeting of the Federation of the Society of Experimental Biology, Atlantic City, New Jersey, April, 1973.

² This work is taken in part from the project report submitted by Kim Goldenberg in partial fulfillment of the requirements for the Master of Science degree in Bioengineering at the Polytechnic Institute of Brooklyn, Brooklyn, New York.

³ Requests for reprints should be sent to Doctor Praissman, Department of Medicine, Nassau County Medical Center, East Meadow, New York 11554.