Venable John D, Wohlschlegel James, McClatchy Daniel B, Park Sung Kyu, Yates John R
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92014, USA.
Anal Chem. 2007 Apr 15;79(8):3056-64. doi: 10.1021/ac062054i. Epub 2007 Mar 17.
The quantitative analysis of complex biological samples has emerged as a key research area in the field of proteomics. Although quantitative proteomic experiments remain challenging, these strategies have been greatly facilitated by the development of newer high-performance mass spectrometers. In this work, we have evaluated the use of the LTQ-Orbitrap, a hybrid mass spectrometer in which a linear ion trap is coupled to an Orbitrap mass analyzer, for quantitative analyses. By analyzing a range of yeast protein standards, we found that the high mass accuracy, high resolution, large ion capacity, and large dynamic range of the LTQ-Orbitrap led to as much as a 4-5-fold improvement in the number and quality of the peptide ratio measurements compared to similar analyses done on the LTQ. We also successfully quantified protein expression differences that occur in metabolically labeled rat synapses during brain development to further demonstrate the suitability of the LTQ-Orbitrap for the comparative analysis of complex tissue samples.
复杂生物样品的定量分析已成为蛋白质组学领域的一个关键研究领域。尽管定量蛋白质组学实验仍然具有挑战性,但更新的高性能质谱仪的发展极大地推动了这些策略。在这项工作中,我们评估了LTQ-Orbitrap(一种线性离子阱与轨道阱质量分析器耦合的混合质谱仪)在定量分析中的应用。通过分析一系列酵母蛋白质标准品,我们发现LTQ-Orbitrap的高质量准确性、高分辨率、大离子容量和大动态范围,与在LTQ上进行的类似分析相比,肽比率测量的数量和质量提高了多达4至5倍。我们还成功地量化了大脑发育过程中代谢标记的大鼠突触中发生的蛋白质表达差异,以进一步证明LTQ-Orbitrap适用于复杂组织样品的比较分析。