Ultra-performance liquid chromatography/mass spectrometry of intact proteins
Ultra-performance liquid chromatography/mass spectrometry of intact proteins
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DOI：
10.1016/j.chroma.2008.03.058
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发表时间：
2008-05-30
影响因子：
4.1
通讯作者：
Croley, Timothy R.
中科院分区：
文献类型：
--
作者：
Everley, Robert A.;Croley, Timothy R.
鉴于超高效液相色谱（UPLC）的许多小分子应用已经发表，我们努力研究UPLC在增强完整蛋白质分离方面的潜力。从典型的使用条件开始，优化柱温和有机溶剂，然后进行HPLC和UPLC的比较。当应用于10种蛋白质标准品的混合物时，优化的方法产生了改进的色谱分辨率，增强的灵敏度和吞吐量增加了三倍。随后的细胞裂解液分析表明，在原始运行时间的1/3下，色谱或质谱数据质量没有妥协。(c) 2008 Elsevier B.V.版权所有
Given that numerous small molecule applications of ultra-performance liquid chromatography (UPLC) have been published, efforts were made to examine the potential of UPLC to enhance the separation of intact proteins. Beginning with typically employed conditions, column temperature and organic solvent were optimized followed by an HPLC vs. UPLC comparison. When applied to a mixture of 10 protein standards, the optimized method yielded improved chromatographic resolution, enhanced sensitivity, and a threefold increase in throughput. Subsequent cell lysate analysis demonstrated no compromise in chromatographic or mass spectral data quality at 1/3 of the original run time. (c) 2008 Elsevier B.V. All rights reserved.