A radiochemical-high-performance liquid chromatographic assay for urate oxidase in human plasma

✍ Scribed by Michael L. Greenberg; Michael S. Hershfield

• Publisher: Elsevier Science
• Year: 1989
• Tongue: English
• Weight: 381 KB
• Volume: 176
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(89)90311-4

✦ Synopsis

Polyethylene

glycol-modified urate oxidase (PEG-uricase) holds promise as a hypouricemic agent for treating gout and as an adjunct to cytolytic therapy of hematologic malignancies. Spectrophotometric assays of urate oxidase are not sensitive enough for pharmacokinetic evaluation of PEG-uricase in clinical trials. We have therefore developed a more sensitive radiochemical-HPLC assay for urate oxidase activity in untreated plasma, in which 14C in urate and in the reaction product, allantoin, is monitored in the uv detector effluent with a flow-through scintillation counter.

The assay is linear with amount of enzyme and time of incubation and can detect <l X lo-' U/ml uricase in plasma. The assay accounts for plasma samples of widely differing urate content.