Fast, Comprehensive Two-Dimensional HPLC Separation of Tryptic Peptides Based on High-Temperature HPLC

详细信息    查看全文

• 作者：Dwight R. Stoll and Peter W. Carr
• 刊名：Journal of the American Chemical Society
• 出版年：2005
• 出版时间：April 13, 2005
• 年：2005
• 卷：127
• 期：14
• 页码：5034 - 5035
• 全文大小：39K
• 年卷期：v.127,no.14(April 13, 2005)
• ISSN：1520-5126

文摘

Two-dimensional HPLC (2D-LC) has recently received considerable attention, and is being used as an alternative to 2D gel electrophoresis in proteomics research. The greatest impediment to the widespread use of 2D-LC is the long analysis time ranging up to days per analysis, making the technique impractical for many jobs. Here we focus on improving the speed of gradient separations since these are typically used as the second dimension in peptide separations by 2D-LC. Specifically we describe high-temperature, ultrafast HPLC conditions, along with the instrument modifications needed to reduce the analysis time of each complete second-dimension gradient separation to tens of seconds. Most importantly, this system is capable of generating a high peak capacity (1350) characteristic of comprehensive 2D-LC in a relatively shorter analysis time (20 min) with a sampling rate sufficient to minimize information loss with simpler instrumentation than currently used; this is equivalent to one unit of peak capacity per second.