ETHANOL FRACTIONATION OF PLASMA AND ELECTROPHORETIC IDENTIFICATION OF INSULIN-BINDING ANTIBODY

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Abstract
The sera of 9 insulin-treated subjects were fractionated with cold ethanol. The fractions were tested for the presence of insulin-binding antibody employing I131 labeled crystalline beef insulin and paper strip electrophoresis. In all cases the antibody was present in fraction I + III (beta and alpha globulins). In 5 cases antibody was also present in fraction II (mainly gamma globulin) but in 3 of these cases refractionation of the same serum revealed antibody to be present only in fraction I + III. The zone of migration of the antibody-insulin complex was in the inter gamma-beta region. The distribution of antibody among the fractions indicates that complexed and uncomplexed insulin-binding antibody have essentially the same mobility. There was no difference, qualitatively, in the recovery of antibody in specific fractions when insulin-I131 was added to whole plasma or serum before or after fractionation.