Preparation of human antibodies to insulin

Insulin was directly coupled and also indirectly coupled by a side chain to Sepharose 4B to form immunoabsorbents for affinity chromatography of insulin antibodies. The two direct derivatives were insulin lysyl and insulin phenylalanyl sepharose. The indirect derivative, insulin carboxyhexyl sepharose, was prepared in two ways: by a carbodiimide procedure and by an N-hydroxysuccinimide ester method. Insulin carboxyhexyl sepharose prepared by the latter method was found to be the most satisfactory material for affinity chromatography and was used to isolate insulin antibodies from human serum.