Department of Laboratory Medicine, Division of Clinical Chemistry, Lund University, Skåne University Hospital, 20502 Malmö, Sweden.
J Proteomics. 2012 Dec 5;76 Spec No.:116-24. doi: 10.1016/j.jprot.2012.05.003. Epub 2012 May 9.
The aim of this study is a novel automated sample-processing concept for future proteomics and clinical research, performing patient studies from resulting blood fractions in various disease areas. Another aim is biobank storage of small sample volumes, where each sample aliquot can be used for a dedicated clinical analysis and end-point measurement in order to preserve sample integrity and value over time.
96 and 384 format sample storage tube systems were utilized for preservation and archiving of clinical patient samples. Automated sample processing and aliquoting were achieved using robotic liquid handling instrumentation, followed by biomarker assay quantitations. Sample workflow was documented and tracked by Nautilus LIMS.
Validation by repetitive processing and analysis confirmed the reliability of automated high density 384 format aliquoting. This high density scaling allows for reproducible aliquoting of 70-μL volumes of blood. Plasma with EDTA, Li-heparin, and citrate, as anti-coagulants, fractioned along with the buffy coat (leukocytes) and the erythrocyte fraction. Large scale processing of 11,000 sample aliquots resulted in a 99.8% process fulfillment.
Our results demonstrate that robust results can be generated from an automated sample processing strategy, isolating plasma, buffy coat, erythrocytes, serum and whole blood, proven by quantitation of 23 common markers used in everyday healthcare around the world. This article is part of a Special Issue entitled: Integrated omics.
本研究的目的是开发一种新颖的自动化样品处理概念,用于未来的蛋白质组学和临床研究,从各种疾病领域的血液馏分中进行患者研究。另一个目的是生物银行储存小样本量,其中每个样本等分可以用于专门的临床分析和终点测量,以保持样本的完整性和随时间的价值。
使用 96 和 384 格式的样品储存管系统保存和存档临床患者样品。使用机器人液体处理仪器进行自动化样品处理和等分,然后进行生物标志物测定定量。通过 Nautilus LIMS 记录和跟踪样品工作流程。
通过重复处理和分析进行验证,证实了自动化高密度 384 格式等分的可靠性。这种高密度扩展允许可重复地等分 70μL 体积的血液。EDTA、Li-肝素和柠檬酸盐作为抗凝剂,与白细胞的血涂片和红细胞部分一起分离。对 11000 个样品等分的大规模处理导致 99.8%的过程完成。
我们的结果表明,从自动化样品处理策略可以产生稳健的结果,通过对全球日常医疗保健中使用的 23 种常见标记物的定量证明,可以分离血浆、血涂片、红细胞、血清和全血。本文是一个特刊的一部分,主题为:综合组学。