Abstract
A derivatized insulin, succinyl amido phenyl-α- d-glycopyranoside insulin (SAPG-insulin) has been synthesized for use in a self-regulating insulin delivery system. The product was a heterogenous mixture of glycosylated insulins differing in their degree and site of substitution. These products were purified and separated using Con A affinity chromatography followed by fast protein liquid chromatography (FPLC) on an anion exchange column. The resultant 4 products were isolated and characterized by end group analysis, sugar group quantitation and isoelectric focusing. In addition, radioimmunoassay cross-reactivity and in vivo biological activity were assessed. The major peak was B-1 phenylalanine monosubstituted SAPG-insulin, which had a potency equivalent to native bovine insulin. This makes the purified fraction, B-1 phenylalanine monosubstituted SAPG-insulin, an ideal candidate for use in the self-regulating insulin delivery system. The method utilized in this work is also potentially useful in the modification of proteins or peptides for enhanced gastrointestinal absorption.
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