优化基于二氧化钛的磷酸肽富集方法以用于与串联质谱联用的自动化多维液相色谱分析
Optimizing TiO2-based phosphopeptide enrichment for automated multidimensional liquid chromatography coupled to tandem mass spectrometry.
作者信息
Cantin Greg T, Shock Teresa R, Park Sung Kyu, Madhani Hiten D, Yates John R
机构信息
Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, SR11, La Jolla, California 92037, USA.
出版信息
Anal Chem. 2007 Jun 15;79(12):4666-73. doi: 10.1021/ac0618730. Epub 2007 May 25.
Abstract
An automated online multidimensional liquid chromatography system coupled to ESI-based tandem mass spectrometry was used to assess the effectiveness of TiO2 in the enrichment of phosphopeptides from tryptic digests of protein mixtures. By monitoring the enrichment of phosphopeptides, an optimized set of loading, wash, and elution conditions were realized for TiO2. A comparison of TiO2 with other resins used for phosphopeptide enrichment, Fe(III)-IMAC and ZrO2, was also carried out using tryptic digests of both simple and moderately complex protein mixtures; where TiO2 was shown to be superior in performance.
摘要
使用与基于电喷雾电离的串联质谱联用的自动化在线多维液相色谱系统，来评估二氧化钛从蛋白质混合物的胰蛋白酶消化物中富集磷酸化肽段的有效性。通过监测磷酸化肽段的富集情况，实现了针对二氧化钛的一组优化的上样、洗涤和洗脱条件。还使用简单和中等复杂程度蛋白质混合物的胰蛋白酶消化物，对二氧化钛与其他用于磷酸化肽段富集的树脂（Fe(III)-IMAC和二氧化锆）进行了比较；结果表明二氧化钛的性能更优。
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