Quantitative Urinary Proteome Analysis for Biomarker Evaluation in Chronic Kidney Disease
Quantitative Urinary Proteome Analysis for Biomarker Evaluation in Chronic Kidney Disease
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DOI：
10.1021/pr800401m
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发表时间：
2009-01-01
影响因子：
4.4
通讯作者：
Metzger, Jochen
中科院分区：
文献类型：
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作者：
Jantos-Siwy, Justyna;Schiffer, Eric;Metzger, Jochen
蛋白质组学方法在其临床适用性方面的限制是缺乏从特定质谱（MS）谱直接推断蛋白质或肽的量的可能性。对于定量的慢性肾脏疾病（CKD）-特异性尿多肽的毛细管电泳-质谱（CE-MS），我们比较了信号强度校准方法的基础上，无论是尿肌酐或稳定同位素标记的合成标记物类似物（绝对定量）与离子计数的基础上，使用高丰度的胶原蛋白片段作为非标记参考（相对定量）。我们的研究结果表明，相对定量的生物标志物排泄的离子计数的基础上参考内源性“管家”肽是足够的尿多肽水平的测定。通过外源性稳定同位素标记的肽标准品计算绝对浓度没有额外的益处。
A limitation of proteomic methods with respect to their clinical applicability is the lack of possibilities to directly deduce the amount of a protein or peptide from a particular mass spectrometry (MS) spectrum. For quantification of chronic kidney disease (CKD)-specific urinary polypeptides in capillary electrophoresis coupled with mass spectrometry (CE-MS), we compared signal intensity calibration methods based on either urinary creatinine or stable isotope labeled synthetic marker analogues (absolute quantification) with those based on ion counting using highly abundant collagen fragments as nonmarker references (relative quantification). Our results indicate that relative quantification of biomarker excretion based on ion counts in reference to endogenous "housekeeping" peptides is sufficient for the determination of urinary polypeptide levels. The calculation of absolute concentrations via exogenous stable isotope-labeled peptide standards is of no additional benefit.