Some quantitative aspects of the labelling of proteins with 125 I by the iodine monochloride method.

Abstract

The labelling of proteins by the iodine monochloride method was studied by using a mathematical model. The equations used were primarily derived from the mass law equation of the isotopic exchange reaction between [(125)I]iodide and iodine monochloride. For convenient application, all equations were programmed into a computing desk-top calculator. To support the validity of the theoretical model, a series of iodinations of insulin were performed under various labelling conditions. The results of these experiments compare well with the theoretically derived values. Deviations from the theoretical values occurring at molar ratios of [(125)I]iodide to iodine monochloride < 0.1 and > 4.0 are explained and suggestions made about how to prevent them. The mathematical model was used to simulate the isotopic exchange, and the iodination reaction under various conditions, to study (a) the influence of the amount of [(125)I]iodide on the amount of [(125)I]iodine monochloride formed, (b) the influence of the specific radioactivity of [(125)I]iodide on the amount of [(125)I]iodine monochloride formed, and (c) the influence of the specific radioactivity of [(125)I]iodide on the number of millicuries needed for labelling to a desired extent.