Department of Chemistry, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4R2.
Anal Chem. 2021 Oct 26;93(42):14042-14047. doi: 10.1021/acs.analchem.1c03549. Epub 2021 Oct 13.
Sodium dodecyl sulfate (SDS) provides numerous benefits for proteome sample preparation. However, the surfactant can be detrimental to downstream mass spectrometry analysis. Although strategies are available to deplete SDS from proteins, each is plagued by unique deficiencies that challenge their utility for high-throughput proteomics. An optimal approach would rapidly and reproducibly achieve less than 10 ppm residual SDS while simultaneously maximizing analyte recovery. Here, we describe improvements to a simple electrokinetic device termed transmembrane electrophoresis, which we previously reported for automated, rapid SDS depletion of proteome samples. Voltage-driven transport of SDS across a molecular weight cutoff membrane is enhanced at higher electric fields, which is herein achieved by integrating an active cooling mechanism to mitigate the impacts of Joule heating. We report 99.9% reduction of SDS (final concentration < 5 ppm) in 5 min. The device is employed in a detergent-based proteomic workflow for analysis of an enriched yeast membrane proteome extract, demonstrating quantitative protein recovery (>98%) and increasing the number of identifications by liquid chromatography-tandem mass spectrometry.
十二烷基硫酸钠（SDS）在蛋白质组样品制备中具有许多优点。然而，这种表面活性剂会对下游的质谱分析造成损害。虽然有多种去除蛋白质中 SDS 的策略，但每种策略都存在独特的缺陷，这限制了它们在高通量蛋白质组学中的应用。一种理想的方法应该能够快速、可重复地将 SDS 的残留量降低到 10ppm 以下，同时最大限度地提高分析物的回收率。在这里，我们描述了对一种简单的电动设备（称为跨膜电泳）的改进，我们之前曾报道过该设备可用于自动、快速去除蛋白质组样品中的 SDS。在更高的电场下，通过分子量截止膜的 SDS 电驱动传输会得到增强，这是通过集成主动冷却机制来减轻焦耳加热的影响来实现的。我们报告在 5 分钟内 SDS 的去除率达到 99.9%（最终浓度<5ppm）。该设备用于基于去污剂的蛋白质组学工作流程，分析富集的酵母膜蛋白质组提取物，证明了定量的蛋白质回收（>98%）和增加了液相色谱-串联质谱的鉴定数量。
