A simple inulin assay for renal clearance determination using an immobilized beta-fructofuranosidase.
Current hospital practice for testing renal function is to use the creatinine clearance test. Inulin clearance, an inherently more accurate procedure, currently is only carried out by specialized laboratories because there is not a simple biochemical assay for inulin, that is, an assay that could be carried out by any laboratory without special facilities. We have developed a simple enzymatic assay system for measuring inulin in plasma and urine. The procedure uses a beta-fructofuranosidase immobilized on Concanavalin A to convert inulin to fructose. Fructose is then measured by measuring the NADH----NAD conversion produced when fructose is converted to sorbitol by the enzyme sorbitol dehydrogenase. Kinetic parameters, binding capacities, and operating conditions for the immobilized beta-fructofuranosidase were determined as well as general operating parameters for the complete assay system. This system offers the potential for replacing the creatinine clearance test as the assay of choice for renal function.
D004800:Enzymes, Immobilized / Q000378:metabolism; D005632:Fructose / Q000378:metabolism; D005919:Glomerular Filtration Rate; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D007444:Inulin / Q000097:blood / Q000652:urine*; D007668:Kidney / Q000502:physiology*; D007064:L-Iditol 2-Dehydrogenase / Q000378:metabolism; D013012:Sorbitol / Q000378:metabolism; D043324:beta-Fructofuranosidase
D004800:Enzymes, Immobilized; D005632:Fructose; D013012:Sorbitol; D007444:Inulin; D007064:L-Iditol 2-Dehydrogenase; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase