Zürbig Petra, Renfrow Matthew B, Schiffer Eric, Novak Jan, Walden Michael, Wittke Stefan, Just Ingo, Pelzing Matthias, Neusüss Christian, Theodorescu Dan, Root Karen E, Ross Mark M, Mischak Harald
Mosaiques Diagnostics & Therapeutics AG, Hannover, Germany.
Electrophoresis. 2006 Jun;27(11):2111-25. doi: 10.1002/elps.200500827.
CE-MS is a successful proteomic platform for the definition of biomarkers in different body fluids. Besides the biomarker defining experimental parameters, CE migration time and molecular weight, especially biomarker's sequence identity is an indispensable cornerstone for deeper insights into the pathophysiological pathways of diseases or for made-to-measure therapeutic drug design. Therefore, this report presents a detailed discussion of different peptide sequencing platforms consisting of high performance separation method either coupled on-line or off-line to different MS/MS devices, such as MALDI-TOF-TOF, ESI-IT, ESI-QTOF and Fourier transform ion cyclotron resonance, for sequencing indicative peptides. This comparison demonstrates the unique feature of CE-MS technology to serve as a reliable basis for the assignment of peptide sequence data obtained using different separation MS/MS methods to the biomarker defining parameters, CE migration time and molecular weight. Discovery of potential biomarkers by CE-MS enables sequence analysis via MS/MS with platform-independent sample separation. This is due to the fact that the number of basic and neutral polar amino acids of biomarkers sequences distinctly correlates with their CE-MS migration time/molecular weight coordinates. This uniqueness facilitates the independent entry of different sequencing platforms for peptide sequencing of CE-MS-defined biomarkers from highly complex mixtures.
毛细管电泳-质谱联用（CE-MS）是一种成功的蛋白质组学平台，可用于鉴定不同体液中的生物标志物。除了用于定义生物标志物的实验参数（CE迁移时间和分子量）外，生物标志物的序列同一性对于深入了解疾病的病理生理途径或进行量身定制的治疗药物设计而言，是不可或缺的基石。因此，本报告详细讨论了不同的肽测序平台，这些平台由与不同的串联质谱（MS/MS）设备（如基质辅助激光解吸电离飞行时间串联质谱仪（MALDI-TOF-TOF）、电喷雾离子阱质谱仪（ESI-IT）、电喷雾四极杆飞行时间质谱仪（ESI-QTOF）和傅里叶变换离子回旋共振质谱仪）在线或离线耦合的高效分离方法组成，用于对指示性肽段进行测序。这种比较展示了CE-MS技术的独特特性，即它能为将使用不同分离MS/MS方法获得的肽序列数据与定义生物标志物的参数（CE迁移时间和分子量）相关联提供可靠依据。通过CE-MS发现潜在生物标志物能够借助与平台无关的样品分离方式，通过MS/MS进行序列分析。这是因为生物标志物序列中的碱性和中性极性氨基酸数量与其CE-MS迁移时间/分子量坐标明显相关。这种独特性便于不同的测序平台独立用于对来自高度复杂混合物的CE-MS定义的生物标志物进行肽测序。
