A modified radioimmunoassay for arylsulfatase A in human serum and urine.
A radioimmunoassay was developed for the determination of arylsulfatase A (EC 3.1.6.1) in human serum and urine. An isoenzyme of arylsulfatase A purified from human urine was used as a standard antigen. The enzyme was radioiodinated with 125I using the Chloramine T method and was stable for about 4 wk. Antibody-bound enzyme was separated from free enzyme by means of a double antibody technique in the presence of polyethylene glycol (PEG). The working range of the method was 0.15-5.0 ng of arylsulfatase A per assay. The within-assay CV was about 8% for both biological fluids and the between-assay CV for serum was 14.1%. Analytical recoveries were 93.2 +/- 9.1% and 97.8 +/- 5.5% for serum and urine, respectively, and the sensitivity was 0.040 ng of arylsulfatase per assay. Serum samples of 50 healthy blood donors were assayed to establish the normal serum level of immunoreactive enzyme, which was found to be 8.3 ng/ml +/- 1.8 ng/ml of serum. Storage of frozen serum was shown to have no significant effect on results obtained using this RIA.
D000328:Adult; D002553:Cerebroside-Sulfatase / Q000032:analysis*; D003110:Colonic Neoplasms / Q000201:enzymology; D006801:Humans; D008297:Male; D008875:Middle Aged; D011863:Radioimmunoassay / Q000379:methods*; D013429:Sulfatases / Q000032:analysis*
D013429:Sulfatases; D002553:Cerebroside-Sulfatase