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- W1855336105 abstract "Release of α-l-fucosidase (AFU; EC 3.2.1.51) from the cells to the extracellular fluid can reflect an inherited or acquired disease involving the lysosome apparatus or a nonspecific lesion leading to cell lysis. Increased activities of AFU have been observed in the sera of patients with diabetes mellitus, alcoholic cirrhosis, and gastric carcinoma (1). Deugnier et al. (2) first proposed that serum AFU activity is a useful marker in the diagnosis of hepatocellular carcinoma (3). Its use as a diagnostic test, however, is limited by the absence of well-defined reference values and the lack of a sensitive and rapid automated enzyme assay.In most cases, AFU activity is determined by a modification of the method of Zielke et al. (4). This method is limited by its long incubation time (30–60 min), which reflects the poor affinity of the enzyme for the colorimetric substrate used (4-nitrophenyl-α-l-fucoside; 4-NPF). Wood (5) observed that specific activities obtained with fluorescent 4-methyl-umbelliferyl-α-l-fucoside (4-MUF) substrate are more than twice those obtained with 4-NPF. However, adaptation of this method as described by Lombardo et al. (6) on the CLA 1500 continuous-flow analyzer (Carlo Erba) leads to a large expense for substrate. Here we describe the optimization of the fluorometric assay and its micromethod adaptation to the Monarch 2000 centrifugal automated analyzer (Instrumentation Laboratory) fitted with a spectrofluorimeter. We further describe the distribution of plasma AFU activity on a healthy French adult population.All chemicals were purchased from Sigma except the 4-methylumbelliferone (4-MU) calibrator, which was obtained from Sebia. The working substrate solution, 0.25 mmol/L 4-MUF, was obtained by dilution of a 5 mmol/L 4-MUF solution (in methanol-water; 1:2, by volume) in an acidic buffer (0.1 mol/L citrate-phosphate buffer, pH 5.0). The working 50 and 100 μmol/L 4-MU calibrator solutions were obtained by dilution …" @default.
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- W1855336105 title "Fluorimetric Measurement of Plasma α-l-Fucosidase Activity with a Centrifugal Analyzer: Reference Values in a Healthy French Adult Population" @default.
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- W1855336105 doi "https://doi.org/10.1093/clinchem/46.4.560f" @default.
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