Targeted data acquisition using nano liquid chromatrography (nano‐LC) coupled mass spectrometry is an emerging approach when there is a need to quantify proteins with high accuracy, sensitivity, and reproducibility. Nevertheless, creating assays meeting all those criteria still remains a laborious task, especially when investigating low abundant proteins and small concentration changes. In this work a targeted data acquisition workflow is developed reducing time and effort to target and investigate key players of metabolic pathways during the process of adipocyte differentiation. This leads to accurate and sensitive quantification of proteins involved in the synthesis of fatty acids, glycerolipids, glycerophospholipids, sphingolipids, the production of energy and reduction equivalents. Additionally low abundant signaling molecules part of the peroxisome proliferator‐activated receptor gamma (PPARγ) and insulin signaling pathway with ≈400 for the insulin receptor substrate and 1100 copies per cell for PPARγ are determined.

中文翻译：

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开发有针对性的蛋白质组学方法，以监测和量化脂肪生成过程中代谢途径的关键参与者的稳健分析。

当需要以高精度，高灵敏度和可重复性对蛋白质进行定量分析时，使用纳米液相色谱（nano-LC）耦合质谱进行目标数据采集是一种新兴方法。尽管如此，创建满足所有这些标准的测定仍然是一项艰巨的任务，尤其是在研究低丰度蛋白质和小浓度变化时。在这项工作中，开发了有针对性的数据采集工作流程，减少了在脂肪细胞分化过程中靶向和研究代谢途径关键参与者的时间和精力。这导致涉及脂肪酸，甘油脂，甘油磷脂，鞘脂，能量产生和还原当量合成的蛋白质的准确和灵敏的定量。