An integrated digital microfluidic chip for multiplexed proteomic sample preparation and analysis by MALDI-MS
An integrated digital microfluidic chip for multiplexed proteomic sample preparation and analysis by MALDI-MS
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DOI：
10.1039/b601954d
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发表时间：
2006-01-01
期刊：
影响因子：
6.1
通讯作者：
Kim, Chang-Jin CJ''
中科院分区：
文献类型：
--
作者：
Moon, Hyejin;Wheeler, Aaron R.;Kim, Chang-Jin CJ''
为了在数字微流体芯片上实现多路复用样品制备，用于高通量基质辅助激光解吸/电离质谱（MALDI-MS），需要集成几种流体功能。其中包括从储层中产生多个液滴和并行的在线样品纯化。在本文中，我们在处理蛋白质溶液和用电解仪（EWOD）启动的标准蛋白质组学试剂中开发了两个关键的新功能，从而导致用于MALDI-MS多路复用样品制备的集成芯片。第一个是一个电压序列，旨在从三个储层中的每个液滴中生成一系列液滴 - 蛋白质组样品，冲洗流体和MALDI试剂。这是第一次使用EWOD在空气中（而不是石油）环境中分配蛋白质组学试剂。第二个是开发的盒装电极图案，以允许将液滴越过干燥的样品斑点，从而使在线样品纯化的过程可鲁棒进行并行处理。结果，在集成的Ewod-Maldi-MS芯片上证明了多个样品液滴的并行处理，这是迈向高通量MALDI-MS的重要一步。直接从集成设备收集的MS结果具有良好的质量，这表明可以在MALDI-MS应用以及其他高通量蛋白质组学应用程序的芯片上自动化样品制备过程。
To realize multiplexed sample preparation on a digital microfluidic chip for high-throughput Matrix Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS), several fluidic functions need to be integrated. These include the generation of multiple droplets from a reservoir and parallel in-line sample purification. In this paper, we develop two critical new functions in handling protein solutions and standard proteomic reagents with electrowetting-on-dielectric (EWOD) actuation, leading to an integrated chip for multiplexed sample preparation for MALDI-MS. The first is a voltage sequence designed to generate a series of droplets from each of the three reservoirs -proteomic sample, rinsing fluid, and MALDI reagents. It is the first time that proteomic reagents have been dispensed using EWOD in an air (as opposed to oil) environment. The second is a box-in-box electrode pattern developed to allow droplet passing over dried sample spots, making the process of in-line sample purification robust for parallel processing. As a result, parallel processing of multiple sample droplets is demonstrated on the integrated EWOD-MALDI-MS chip, an important step towards high-throughput MALDI-MS. The MS results, collected directly from the integrated devices, are of good quality, suggesting that the tedious process of sample preparation can be automated on-chip for MALDI-MS applications as well as other high-throughput proteomics applications.