游离DNA血液采集管适用于临床蛋白质组学：结直肠癌的实例验证
Cell-Free DNA Blood Collection Tubes Are Appropriate for Clinical Proteomics: A Demonstration in Colorectal Cancer.
作者信息
Almazi Juhura G, Pockney Peter, Gedye Craig, Smith Nathan D, Hondermarck Hubert, Verrills Nicole M, Dun Matthew D
机构信息
Priority Research Centre for Cancer Research, Innovation and Translation, Hunter Medical Research Institute Cancer Research Program, School of Biomedical Sciences and Pharmacy, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.
School of Medicine and Public Health, Faculty of Health and Medicine, University of Newcastle, Callaghan, NSW, Australia.
出版信息
Proteomics Clin Appl. 2018 May;12(3):e1700121. doi: 10.1002/prca.201700121. Epub 2018 Mar 30.
BACKGROUND
Optimized blood collection tubes (BCT) have been developed to expand the utility of plasma cell-free DNA (cfDNA) and are in clinical use. The appropriateness of plasma collected and stored in these tubes for proteomic analysis is unknown.
METHODS
Paired blood samples were collected in BCT and traditional K3EDTA (EDTA) tubes from healthy controls and from colorectal cancer (CRC) patients before and after surgery, and stored for between 45 min and 48 h at room temperature. Plasma proteins were analyzed following high-abundant plasma protein depletion in quantitative discovery and targeted proteomics by liquid chromatography tandem-mass spectrometry (LC-MS/MS).
RESULTS
BCT reduced cellular protein contamination in healthy controls over time, and increased the number of high confident low-abundant protein identifications in CRC blood samples compared to matched samples collected in EDTA tubes. The known CRC plasma protein biomarker, carcinoembryonic antigen (CEA), showed elevated levels across patients pre-operatively when collected and stored in BCT compared to EDTA tubes. Emerging CRC biomarkers, Dickkopf-3 (DKK3) and Gelsolin (GSN), showed elevated levels pre-operatively when collected in BCT.
CONCLUSIONS
Optimized BCT are appropriate for low-abundant plasma protein analysis and can be used with confidence for clinical proteomics.
背景
已开发出优化的采血管（BCT）以扩大血浆游离DNA（cfDNA）的应用范围，且这些采血管已投入临床使用。目前尚不清楚在这些采血管中采集和储存的血浆用于蛋白质组学分析是否合适。
方法
从健康对照者以及结直肠癌（CRC）患者手术前后分别采集配对的血液样本，分别置于BCT管和传统的K3EDTA（EDTA）管中，并在室温下储存45分钟至48小时。在通过液相色谱串联质谱（LC-MS/MS）进行的定量发现和靶向蛋白质组学中，在去除高丰度血浆蛋白后对血浆蛋白进行分析。
结果
随着时间的推移，BCT减少了健康对照者血浆中细胞蛋白的污染，并且与在EDTA管中采集的匹配样本相比，增加了CRC血样中高可信度低丰度蛋白的鉴定数量。已知的CRC血浆蛋白生物标志物癌胚抗原（CEA），与EDTA管相比，当在BCT中采集和储存时，术前患者的CEA水平升高。新出现的CRC生物标志物Dickkopf-3（DKK3）和凝溶胶蛋白（GSN），在BCT中采集时术前水平升高。
结论
优化的BCT适用于低丰度血浆蛋白分析，可放心用于临床蛋白质组学。
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