Department of Chemistry, University of Tennessee, Knoxville, TN 37996-1600, USA.
J Chromatogr A. 2010 Jul 16;1217(29):4779-83. doi: 10.1016/j.chroma.2010.05.015. Epub 2010 May 20.
Liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) allows the acquisition of a vast amount of analytical data in the aim of identifying peptides and proteins. Difficulties arise when attempting to identify proteins from the results of analyses of their peptide digests. We investigated possible quantitative correlations between the peak capacity achieved in chromatographic analyses and the protein sequence coverage. For this purpose, we used high performance liquid chromatography (HPLC) columns packed with fully and superficially porous (shell) particles of average sizes ranging from 2.6 to 15 microm in diameter. We found that the sequence coverage of bovine serum albumin obtained with a 10-min gradient elution run on a column packed with 2.6 microm shell particles is greater than that provided by 40-min gradients run on columns packed with 15 or 10 microm particles and equal to those yielded by the same 40-min gradient run on columns packed with 3 and 5 microm particles.
液相色谱-质谱联用/质谱（LC-MS/MS）可以获取大量分析数据，旨在鉴定肽和蛋白质。然而，当尝试根据肽消化物的分析结果鉴定蛋白质时，会遇到困难。我们研究了色谱分析中达到的峰容量与蛋白质序列覆盖率之间可能存在的定量相关性。为此，我们使用了填充有全多孔（壳）和部分多孔（壳）颗粒的高效液相色谱（HPLC）柱，平均粒径范围从 2.6 到 15 微米。我们发现，用 2.6 微米壳颗粒填充的色谱柱进行 10 分钟梯度洗脱得到的牛血清白蛋白的序列覆盖率大于用 15 或 10 微米颗粒填充的色谱柱进行 40 分钟梯度洗脱得到的序列覆盖率，与用 3 和 5 微米颗粒填充的色谱柱进行相同的 40 分钟梯度洗脱得到的序列覆盖率相等。
