Végvári Akos, Magnusson Mattias, Wallman Lars, Ekström Simon, Bolmsjö Gunnar, Nilsson Johan, Miliotis Tasso, Ostling Jörgen, Kjellström Sven, Ottervald Jan, Franzén Bo, Hultberg Hans, Marko-Varga György, Laurell Thomas
Department of Electrical Measurements, Lund University, Lund, Sweden.
Electrophoresis. 2008 Jun;29(12):2696-705. doi: 10.1002/elps.200700816.
As much attention has devoted to the proteome research during the last few years, biomarker discovery has become an increasingly hot area, potentially enabling the development of new assays for diagnosis and prognosis of severe diseases. This is the field of research interest where efforts originating from both academic and industrial groups should jointly work on solutions. In this paper, we would like to demonstrate the fruitful combination of both research domains where the scientific crossroads sprout fresh ideas from the basic research domain and how these are refined and tethered to industrial standards. We will present an approach that is based on novel microfluidic devices, utilizing their benefits in processing small-volume samples. Our biomarker discovery strategy, built around this platform, involves optimized samples processing (based on SPE and sample enrichment) and fast MALDI-MS readout. The identification of novel biomarkers at low-abundance level has been achieved by the utilization of a miniaturized sample handling platform, which offers clean-up and enrichment of proteins in one step. Complete automation has been realized in the form of a unique robotic instrumentation that is able to extract and transfer 96 samples onto standard MALDI target plates with high throughput. The developed platform was operated with a 60 sample turnaround per hour allowing sensitivities in femtomol regions of medium- and low-abundant target proteins from clinical studies on samples of multiple sclerosis and gastroesophageal reflux disease. Several proteins have been identified as new biomarkers from cerebrospinal fluid and esophagus epithelial cells.
在过去几年里，蛋白质组研究受到了广泛关注，生物标志物的发现已成为一个日益热门的领域，有可能推动开发用于严重疾病诊断和预后的新检测方法。这是一个研究兴趣领域，学术团体和产业团体的努力应共同致力于解决方案。在本文中，我们希望展示这两个研究领域的卓有成效的结合，即科学的交叉点如何从基础研究领域萌生出新想法，以及这些想法如何被完善并与行业标准相结合。我们将介绍一种基于新型微流控设备的方法，利用其在处理小体积样品方面的优势。我们围绕这个平台构建的生物标志物发现策略，包括优化样品处理（基于固相萃取和样品富集）和快速基质辅助激光解吸/电离质谱读出。通过利用小型化样品处理平台，在低丰度水平上实现了新型生物标志物的鉴定，该平台能够一步完成蛋白质的净化和富集。以一种独特的机器人仪器的形式实现了完全自动化，该仪器能够以高通量将96个样品提取并转移到标准基质辅助激光解吸/电离靶板上。所开发的平台每小时可处理60个样品，对于来自多发性硬化症和胃食管反流病临床研究的中低丰度靶蛋白，在飞摩尔区域具有灵敏度。已经从脑脊液和食管上皮细胞中鉴定出几种蛋白质作为新的生物标志物。
