Preparation of [3-(2-Phyridyldithio)propionoyl]insulins and Horeseradish Peroxidase-Insulin Conjugates by High-Performance Liquid Chromatographic Separation

Abstract

Five [3-(2-pyridyldithio)propionoyl]insulins (PDP-insulins) were separated preparaticely from the reaction mixture of porcine insulin with N-succinimidyl 3-(2-phyidyldithio)propionate by means of anion-exchange high-preformance liquid chromatography on a TSK gel DEAE-SW column. Horseradich peroxidase (HRP)-insulin conjugates (Gly^Al-HRP-insulin, Lys^B29-HRP-insulin and Gly^Al, Lys^B29-diHRP-insulin) were prepared by the reaction of thiolated HRP and the corresponding PDP-insulins and purified by gel-permeation high-performance liquid chromatography on a TSK gel G3000SW column.