Boley Danielle A, Zhang Zhenbin, Dovichi Norman J
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, 46556, USA.
Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, 46556, USA.
J Chromatogr A. 2017 Nov 10;1523:123-126. doi: 10.1016/j.chroma.2017.07.022. Epub 2017 Jul 8.
While capillary zone electrophoresis (CZE) provides dramatically improved numbers of peptide identifications compared with reversed-phase chromatography for bottom-up proteomics of mass limited samples, CZE inevitably produces lower numbers of peptide identifications than RPLC for larger samples. One reason for this poorer performance is the dead time between injection of samples and subsequent appearance of the fastest moving component. This dead time is typically 25% of the separation window in CZE, but is only 5% of the separation window in gradient elution RPLC. This dead time can be eliminated in CZE by use of a multisegment injection mode where a series of samples is analyzed by injecting each sample while the preceding sample is still being separated. In this paper, we demonstrate that capillary zone electrophoresis employing sequential injections can produce a doubling in peptide identification rate with no degradation in separation efficiency.
虽然对于质量有限的样品进行自下而上的蛋白质组学分析时，与反相色谱法相比，毛细管区带电泳（CZE）能显著提高肽段鉴定的数量，但对于较大的样品，CZE不可避免地比反相液相色谱（RPLC）产生的肽段鉴定数量更少。性能较差的一个原因是样品进样与随后最快移动组分出现之间的死时间。在CZE中，这个死时间通常是分离窗口的25%，但在梯度洗脱RPLC中仅为分离窗口的5%。通过使用多段进样模式可以消除CZE中的这个死时间，即在前面的样品仍在分离时注入每个样品，从而对一系列样品进行分析。在本文中，我们证明采用顺序进样的毛细管区带电泳可以使肽段鉴定率提高一倍，而分离效率不会降低。
