Metzger Jochen, Luppa Peter B, Good David M, Mischak Harald
Mosaiques Diagnostics and Terapeutics AG, Mellendorfer Str. 7-9, Hannover 30625, Germany.
Crit Rev Clin Lab Sci. 2009;46(3):129-52. doi: 10.1080/10408360902805261.
Single biomarker detection is common in clinical laboratories due to the currently available method spectrum. For various diseases, however, no specific single biomarker could be identified. A strategy to overcome this diagnostic void is to shift from single analyte detection to multiplexed biomarker profiling. Mass spectrometric methods were employed for biomarker discovery in body fluids. The enormous complexity of biofluidic proteome compartments implies upstream fractionation. For this reason, mass spectrometry (MS) was coupled to two-dimensional gel electrophoresis, liquid chromatography, surface-enhanced laser desorption/ionization, or capillary electrophoresis (CE). Differences in performance and operating characteristics make them differentially suited for routine laboratory applications. Progress in the field of clinical proteomics relies not only on the use of an adequate technological platform, but also on a fast and efficient proteomic workflow including standardized sample preparation, proteomic data processing, statistical validation of biomarker selection, and sample classification. Based on CE-MS analysis, we describe how proteomic technology can be implemented in a clinical laboratory environment. In the last part of this review, we give an overview of CE-MS-based clinical studies and present information on identity and biological significance of the identified peptide biomarkers providing evidence of disease-induced changes in proteolytic processing and posttranslational modification.
由于目前可用的方法范围，单一生物标志物检测在临床实验室中很常见。然而，对于各种疾病，尚未发现特定的单一生物标志物。克服这一诊断空白的策略是从单一分析物检测转向多重生物标志物分析。质谱方法被用于在体液中发现生物标志物。生物流体蛋白质组区室的巨大复杂性意味着需要进行上游分级分离。因此，质谱（MS）与二维凝胶电泳、液相色谱、表面增强激光解吸/电离或毛细管电泳（CE）联用。性能和操作特性的差异使它们在常规实验室应用中的适用性有所不同。临床蛋白质组学领域的进展不仅依赖于使用适当的技术平台，还依赖于快速高效的蛋白质组学工作流程，包括标准化的样品制备、蛋白质组学数据处理、生物标志物选择的统计验证和样品分类。基于毛细管电泳-质谱分析，我们描述了蛋白质组学技术如何在临床实验室环境中实施。在本综述的最后部分，我们概述了基于毛细管电泳-质谱的临床研究，并提供了已鉴定的肽生物标志物的身份和生物学意义的信息，这些信息证明了疾病引起的蛋白水解加工和翻译后修饰的变化。
