Development of a method for early diagnosis of diabetic of complications by detecting glycation reaction intermediates.

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Glycation occurs during normal aging and at accelerated rates in diabetes and is involved in structural and functional alterations of proteins and other cellular components. In terms of diabetic complications, the glycation hypothesis and the poiyol pathway hypothesis are usually discussed independently. An antibody was raised against fructated lysine in proteins by immunizing fructated lysine-conjugated ovalbumin in rabbits. The affinity purified antibody specifically recognized proteins incubated with fructose, but not with glucose. When bovine serum albumin was incubated with various concentrations of fructose, the reactivity of thee antibody increased in a dose- and time-dependent manner. When soluble proteins prepared from either normal or streptozotocin-induced diabetic rat eyes were analyzed by ELISA using this antibody, an increase in the reactive components was observed as a function of aging as well as under diabetic conditions. Immunoblot analysis showed that lens crystallin reacted highly with this antibody. Since fructose is largely biosynthesized through the polyol pathway, which is enhanced under diabetic conditions and lens is known to have a high activity of this pathway, this antibody is capable of recognizing fructated proteins in viva. Thus this antibody represents a potentially useful tool for investigating two major issues which appear to be involved in diabetic complications, namely, the glycation reaction and the polyol pathway.