Hardouin Julie, Duchateau Magalie, Joubert-Caron Raymonde, Caron Michel
Laboratory of Protein Biochemistry and Proteomics, UMR CNRS 7033 (BioMoCeTi), UFR SMBH, Paris13 University, 93017 Bobigny cedex, France.
Rapid Commun Mass Spectrom. 2006;20(21):3236-44. doi: 10.1002/rcm.2725.
Nanoflow liquid chromatography/mass spectrometry (nanoLC/MS) has become a current tool in proteomics applications increasingly used in the search for new biomarkers. A new integrated microfluidic device (HPLC-Chip), coupled to ion trap mass spectrometry (ITMS), appears as an innovative and robust tool for improving the identifications commonly performed by nanoLC/MS/MS. We tested this device for the identification of proteins obtained from two-dimensional gel electrophoresis or chromatography. The chip allows the measurement of reproducible retention times that, in association with m/z ratios, was found useful for identifying peptide sequences without ambiguity. A sensitivity increase of a factor of at least 5-fold is obtained compared to the results obtained previously in our laboratory by conventional nanoLC/MS/MS on the same ion trap. We conclude that this recently available microfluidic device can be a valuable tool during biomarker discovery programs, particularly identifying low-abundance proteins.
纳流液相色谱/质谱联用技术(nanoLC/MS)已成为蛋白质组学应用中的一种常用工具,越来越多地用于寻找新的生物标志物。一种新型的集成微流控装置(HPLC芯片)与离子阱质谱(ITMS)联用,成为一种创新且强大的工具,可改进通常由nanoLC/MS/MS进行的鉴定工作。我们测试了该装置用于鉴定从二维凝胶电泳或色谱法获得的蛋白质。该芯片能够测量可重复的保留时间,结合质荷比,发现其有助于明确无误地鉴定肽序列。与我们实验室之前在同一离子阱上通过传统的nanoLC/MS/MS获得的结果相比,灵敏度提高了至少5倍。我们得出结论,这种最近可用的微流控装置在生物标志物发现计划中可能是一种有价值的工具,特别是在鉴定低丰度蛋白质方面。