Integrated sample preparation and MALDI mass spectrometry on a microfluidic compact disk
Integrated sample preparation and MALDI mass spectrometry on a microfluidic compact disk
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DOI：
10.1021/ac030194b
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发表时间：
2004-01-15
影响因子：
7.4
通讯作者：
Bergman, T
中科院分区：
文献类型：
--
作者：
Gustafsson, M;Hirschberg, D;Bergman, T
描述了在光盘(CD)上结合基质辅助激光解吸/电离(MALDI)质谱仪的高通量蛋白质消化的微流控加工。离心力使液体通过多个微结构，每个结构包含一个10毫升的反相色谱柱。CD可以并行制备96个样品，体积从一微升到几微升不等。消化中的多肽被浓缩、脱盐，然后使用含有MALDI基质的溶剂从柱上直接洗脱到CD上的MALDI靶区(200×400微米)。结晶后，将CD插入MALDI仪器，以进行多肽质量指纹图谱和数据库鉴定，其灵敏度可降至200-amol水平。检测到低至50-amol水平的蛋白水解肽。CD技术在蛋白质识别方面的成功率大约是C(18)ZipTips和标准MALDI钢靶的两倍。使用机器人操作CD，将样品和试剂从微容器转移到一次性CD上的加工入口，并旋转以控制液体通过微结构的移动。
High-throughput microfluidic processing of protein digests integrated with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry on a compact disk (CD) is described. Centrifugal force moves liquid through multiple microstructures, each containing a 10-nL reversed-phase chromatography column. The CD enables parallel preparation of 96 samples with volumes ranging from one to several microliters. The peptides in the digests are concentrated, desalted, and subsequently eluted from the columns directly into MALDI target areas (200 x 400 mum) on the CD using a solvent containing the MALDI matrix. After crystallization, the CD is inserted into the MALDI instrument for peptide mass fingerprinting and database identification at a routine sensitivity down to the 200-amol level. Detection of proteolytic peptides down to the 50-amol level is demonstrated. The success rate of the CD technology in protein identification is about twice that of the C(18)ZipTips and standard MALDI steel targets. The CDs are operated using robotics to transfer samples and reagents from microcontainers to the processing inlets on the disposable CD and spinning to control the movement of liquid through the microstructures.