Preparation of Fluorescence Labeled Insulins, Sulfobenzoxadiazolyl-insulins, for Fluorescence Immunoassay

Abstract

The preparation of insulins labeled with a fluorescent moiety at a definite position (Gly(A1) or Lys(B29)) on the molecule is described. Gly(A1)- and Lys(B29)-S-acetylthioglycoloyl-insulins and Gly(A1)-2(or 3)-acetylmercapto-3-carboxypropanoyl-insulin were deacetylated and then reacted with 7-chloro-4-sulfobenzoxadiazole, a fluorogenic reagent, to afford the corresponding fluorescence labeled insulins. The preparative separation of the fluorescence labeled insulins from the insulin derivatives that remained unreacted was carried out by anion-exchange high-performance liquid chromatography on a TSKgel DEAE-2SW column.