NONRADIOACTIVE IMMUNOQUANTIFICATION OF ALPHA-L-FUCOSIDASE PROTEIN IN HUMAN COLON TISSUES

alpha-L-Fucosidase is a glycosidase involved in the degradation of fuc oglycoconjugates and has a diagnostic significance because it has been described to be altered in several known diseases, However, in vitro studies on enzymatic activities may not reflect the real protein level s in tissues, This paper describes a simple method to quantify alpha-L -fucosidase protein levels in human crude extracts, combining the slot -blot technique and a nonradioactive immunoassay. Taking advantage of the similarities in different mammalian fucosidases, a polyclonal anti serum was raised against commercial purified alpha-L-fucosidase from b ovine kidney that cross-reacted with the human colon enzyme. The metho d is able to detect as little as 0.75 ng alpha-L-fucosidase. To illust rate the direct application of this technique, we analysed and quantif ied alpha-L-fucosidase protein levels in 18 human colon crude samples, This technique could prove useful in clinical pathology, allowing fas t and accurate measurement of alpha-L-fucosidase in crude extracts.