Sobhani Kimia
Department of Clinical Pathology, Cleveland Clinic, Cleveland, OH, USA.
Methods Mol Biol. 2010;641:325-46. doi: 10.1007/978-1-60761-711-2_18.
The identities and abundance levels of proteins excreted in urine are not only key indicators of diseases associated with renal function but are also indicators of the overall health of individuals. Urine specimens are readily available and provide a noninvasive means to assess and diagnose many disease states. Proteins in urine originate from two sources: the ultrafiltrate of plasma, and those that are shed from the urinary tract. The protein concentration in urine excreted from a normal adult is approximately 150 mg/day, and is typically not greater than 10 mg/100 mL in any single specimen. Following precipitation, concentration, and fractionation methods, proteins of interest from urine samples can be separated, identified, and quantified. One of the most commonly used techniques in the field of urine proteomics is gel electrophoresis followed by identification with mass spectrometry and protein database search algorithms. In this chapter, two-dimensional gel electrophoresis (2-DE) will be discussed, along with less frequently applied techniques, such as isotope coded affinity tags (ICAT) and capillary electrophoresis (CE). Publications discussing the application of these techniques to urine proteomic analyses of healthy individuals and urinary disease biomarker discovery will also be summarized.
尿液中排出的蛋白质的种类和丰度水平不仅是与肾功能相关疾病的关键指标，也是个体整体健康状况的指标。尿液标本易于获取，为评估和诊断多种疾病状态提供了一种非侵入性手段。尿液中的蛋白质有两个来源：血浆超滤液和尿路脱落的蛋白质。正常成年人尿液中排出的蛋白质浓度约为150毫克/天，在任何单个标本中通常不超过10毫克/100毫升。经过沉淀、浓缩和分级分离方法后，可以对尿液样本中感兴趣的蛋白质进行分离、鉴定和定量。尿液蛋白质组学领域最常用的技术之一是凝胶电泳，随后通过质谱和蛋白质数据库搜索算法进行鉴定。在本章中，将讨论二维凝胶电泳（2-DE）以及较少应用的技术，如同位素编码亲和标签（ICAT）和毛细管电泳（CE）。还将总结讨论这些技术在健康个体尿液蛋白质组分析和泌尿系统疾病生物标志物发现中的应用的出版物。
