An Integrated Platform for Isolation, Processing, and Mass Spectrometry-based Proteomic Profiling of Rare Cells in Whole Blood
An Integrated Platform for Isolation, Processing, and Mass Spectrometry-based Proteomic Profiling of Rare Cells in Whole Blood
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DOI：
10.1074/mcp.m114.045724
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发表时间：
2015-06-01
影响因子：
7
通讯作者：
Ivanov, Alexander R.
中科院分区：
文献类型：
--
作者：
Li, Siyang;Plouffe, Brian D.;Ivanov, Alexander R.
生物体液和组织中稀有细胞（例如循环肿瘤和干细胞）的分离和分子表征在临床诊断和个性化医学中具有显著的潜力。目前的工作描述了一个集成的平台，样品采购，制备和分析的血液中的稀有细胞的深度蛋白质组学分析。微流控磁泳分离的目标细胞掺入到1毫升的血液中，在1000-2000个细胞/毫升的水平，然后通过聚焦声学辅助样品制备已耦合与一维PLOT-LC-MS方法。所得到的zeptomole检测灵敏度使得能够在每次分析仅注射相当于100-200个细胞的情况下鉴定类似于4000种蛋白质。通过对第一个加入全血作为模型系统的MCF-7细胞以及志愿者全血中的两个CD 133+内皮祖细胞和造血细胞进行分离和定量蛋白质组学分析，显示了有限体积生理液体中稀有细胞的特征。
Isolation and molecular characterization of rare cells (e.g. circulating tumor and stem cells) within biological fluids and tissues has significant potential in clinical diagnostics and personalized medicine. The present work describes an integrated platform of sample procurement, preparation, and analysis for deep proteomic profiling of rare cells in blood. Microfluidic magnetophoretic isolation of target cells spiked into 1 ml of blood at the level of 1000-2000 cells/ml, followed by focused acoustics-assisted sample preparation has been coupled with one-dimensional PLOT-LC-MS methodology. The resulting zeptomole detection sensitivity enabled identification of similar to 4000 proteins with injection of the equivalent of only 100-200 cells per analysis. The characterization of rare cells in limited volumes of physiological fluids is shown by the isolation and quantitative proteomic profiling of first MCF-7 cells spiked into whole blood as a model system and then two CD133+endothelial progenitor and hematopoietic cells in whole blood from volunteers.