Determination of uromodulin in human urine: influence of storage and processing
Determination of uromodulin in human urine: influence of storage and processing
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DOI：
10.1093/ndt/gft345
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发表时间：
2014-01-01
影响因子：
6.1
通讯作者：
Devuyst, Olivier
中科院分区：
文献类型：
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作者：
Youhanna, Sonia;Weber, Julien;Devuyst, Olivier
尿调蛋白（TammHorsfall蛋白）是在生理条件下尿液中排泄的最丰富的蛋白质。它仅在肾脏中产生，并通过蛋白水解裂解分泌到尿液中。UMOD（编码尿调节蛋白的基因）参与罕见的常染色体显性遗传疾病，其与慢性肾脏疾病风险的全基因组关联表明尿液中尿调节蛋白的水平可能代表肾功能的关键生物标志物。尿调节素结构复杂，具有多个二硫键和典型的胞外蛋白结构域，迄今为止，尿调节素在人体尿液中的稳定性及其测定条件尚未得到系统研究，结果不一致。在这项研究中，我们使用了一个强大的，内部ELISA来表征采样和储存条件，以提供尿液中尿调节素的可靠剂量。人尿液中尿调节素的水平受到离心和涡旋以及储存条件和持续时间的显著影响。这些结果验证了一个简单，低成本的ELISA，并记录了用于测量人尿中尿调节素的最佳处理和储存条件。
Uromodulin (TammHorsfall protein) is the most abundant protein excreted in the urine under physiological conditions. It is exclusively produced in the kidney and secreted into the urine via proteolytic cleavage. The involvement of UMOD, the gene that encodes uromodulin, in rare autosomal dominant diseases, and its robust genome-wide association with the risk of chronic kidney disease suggest that the level of uromodulin in urine could represent a critical biomarker for kidney function. The structure of uromodulin is complex, with multiple disulfide bonds and typical domains of extracellular proteins.Thus far, the conditions influencing stability and measurement of uromodulin in human urine have not been systematically investigated, giving inconsistent results. In this study, we used a robust, in-house ELISA to characterize the conditions of sampling and storage necessary to provide a faithful dosage of uromodulin in the urine.The levels of uromodulin in human urine were significantly affected by centrifugation and vortexing, as well as by the conditions and duration of storage.These results validate a simple, low-cost ELISA and document the optimal conditions of processing and storage for measuring uromodulin in human urine.