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Appl Environ Microbiol. 1974 December; 28(6): 935-942
Copyright © 1975 American Society for Microbiology. All Rights Reserved.

Criteria for Preparing, Evaluating, and Standardizing Iodinated Globulins for Radioimmunoassay Procedures

Center for Disease Control, Atlanta, Georgia 30333

ABSTRACT

Procedures were examined for labeling immune globulins with radioactive iodine using chloramine-T as the oxidizing agent. The chloramine-T method was critically evaluated to establish the optimal conditions for preparing iodinated globulins with high specific radioactivities without impairing their immunospecificities for use in in vitro radioimmunoassays. The results showed that the use of 100 µg of chloramine-T per ml, 500 to 1,000 µCi of Na ¹²⁵I per mg of protein, and a 10-min oxidation reaction time produced globulins of both high specific radioactivities and immunospecificities. Criteria were established for evaluating and determining optimal concentrations of iodine-labeled globulin for use in radioimmunoassays. The results of this investigation indicated that the amount of labeled indicator globulin used in radioimmunoassays should be based upon protein concentration rather than radioactivity.