A RADIOCHEMICAL HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ASSAY FOR URATE OXIDASE IN HUMAN-PLASMA
A RADIOCHEMICAL HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ASSAY FOR URATE OXIDASE IN HUMAN-PLASMA
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DOI：
10.1016/0003-2697(89)90311-4
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发表时间：
1989-02-01
影响因子：
2.9
通讯作者：
HERSHFIELD, MS
中科院分区：
文献类型：
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作者：
GREENBERG, ML;HERSHFIELD, MS
聚乙二醇修饰的尿酸氧化酶（PEG-尿酸酶）有望作为治疗痛风的降尿酸药物和血液恶性肿瘤细胞溶解治疗的辅助药物。尿酸氧化酶的分光光度测定法对于临床试验中PEG-尿酸酶的药代动力学评价不够灵敏。因此，我们开发了一种更灵敏的放射化学-HPLC法测定未处理血浆中的尿酸氧化酶活性，其中尿酸盐和反应产物尿囊素中的14 C在紫外检测器流出液中用流通闪烁计数器进行监测。该测定与酶的量和温育时间呈线性关系，并且可以检测< 1 × 10 - 4。10-5 U/ml血浆尿酸酶。该测定法解释了尿酸盐含量差异很大的血浆样品。
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 in linear with amount of enzyme and time of incubation and can detect < 1 .times. 10-5 U/ml uricase in plasma. The assay accounts for plasma samples of widely differing urate content.