THE PRODUCTION AND IDENTIFICATION OF ANTIBODIES TO INSULIN AND THEIR USE IN ASSAYING INSULIN 1

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Abstract
The intravenous injection of alum-precipitated insulin into rabbits leads to the appearance of antibodies. These antibodies can be detected by their ability to agglutinate insulin-sensitized erythrocytes. These erythrocytes are prepared by conjugating insulin to the red cells by treatment with bis-diazotized benzidine. By this method antibodies to insulin were demonstrated in human diabetics who were resistant to insulin. Antibodies were not demonstrable in a number of diabetics who were not resistant to insulin. When complement was added to the above antigen-antibody system a specific lysis of the insulin-sensitized red cells occurred. This specific immune lysis was inhibited when insulin in solution was preincubated with the antibody before addition of the insulin-sensitized erythrocytes. The degree of inhibition varies with the amount of insulin added. Therefore, this method is applicable to the quantitative determination of insulin. Insulin (0.1 [mu]g) can thereby be detected.

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