Institute of Physiology, Zurich Center for Integrative Human Physiology, University of Zurich, Zurich, Switzerland.
Nephrol Dial Transplant. 2014 Jan;29(1):136-45. doi: 10.1093/ndt/gft345. Epub 2013 Oct 3.
Uromodulin (Tamm-Horsfall 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.
尿调蛋白（Tamm-Horsfall 蛋白）是生理条件下尿液中排泄量最多的蛋白质。它仅在肾脏中产生，并通过蛋白水解切割分泌到尿液中。编码尿调蛋白的 UMOD 基因参与罕见的常染色体显性疾病，并且其与慢性肾脏病风险的全基因组关联表明，尿液中尿调蛋白的水平可能代表肾功能的关键生物标志物。尿调蛋白的结构复杂，具有多个二硫键和典型的细胞外蛋白结构域。
到目前为止，尚未系统研究影响人尿液中尿调蛋白稳定性和测量的条件，导致结果不一致。在这项研究中，我们使用了一种稳健的内部 ELISA 来描述采样和储存条件，以提供尿液中尿调蛋白的真实剂量。
尿调蛋白在人尿中的水平受到离心和涡旋以及储存条件和时间的显著影响。
这些结果验证了一种简单、低成本的 ELISA，并记录了测量人尿中尿调蛋白的最佳处理和储存条件。
