Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
Proteomics. 2012 Nov;12(21):3138-42. doi: 10.1002/pmic.201200168. Epub 2012 Sep 24.
While the use of detergents is necessary for a variety of protein isolation preparation protocols, they are not compatible with mass spectral analysis due to ion suppression and adduct formation. This manuscript describes optimization of detergent removal, using commercially available SDS depletion spin columns containing an affinity resin, providing for both increased protein recovery and thorough SDS removal. Ion mobility spectrometry coupled with mass spectrometry (IMS-MS) allowed for a concurrent analysis of both analyte and detergent. In the case of both proteins and peptides, higher detergent concentrations than previously reported provided an increase of sample recovery; however there was a limit as SDS was detected by IMS-MS at higher levels of SDS indicating incomplete detergent depletion. The results also suggest that optimal conditions for SDS removal are dependent on the sample concentration. Overall, this study provides a useful guide for proteomic studies where SDS is required for efficient sample preparation.
虽然各种蛋白质分离制备方案都需要使用清洁剂,但由于离子抑制和加合物形成,它们与质谱分析不兼容。本文描述了使用市售的含有亲和树脂的 SDS 耗尽旋转柱进行清洁剂去除的优化,这为提高蛋白质回收率和彻底去除 SDS 提供了可能。离子淌度质谱联用(IMS-MS)允许同时分析分析物和清洁剂。对于蛋白质和肽,与之前报道的更高的清洁剂浓度相比,提供了更高的样品回收率;然而,存在一个限制,因为 IMS-MS 检测到更高水平的 SDS 表明 SDS 去除不完全。结果还表明,SDS 去除的最佳条件取决于样品浓度。总的来说,这项研究为需要 SDS 进行有效样品制备的蛋白质组学研究提供了有用的指南。