State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing, China; Beijing Proteome Research Center, Beijing, China; International Academy of Phronesis Medicine, Guangzhou, Guangdong, China; The π-Hub Infrastructure, Guangzhou, Guangdong, China.
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing, China; Guangzhou Laboratory, Guangzhou, Guangdong, China.
Mol Cell Proteomics. 2024 Oct;23(10):100837. doi: 10.1016/j.mcpro.2024.100837. Epub 2024 Sep 6.
Universal sample preparation for proteomic analysis that enables unbiased protein manipulation, flexible reagent use, and low protein loss is required to ensure the highest sensitivity of downstream liquid chromatography-mass spectrometry (LC-MS) analysis. To address these needs, we developed a ZnCl precipitation-assisted sample preparation method (ZASP) that depletes harsh detergents and impurities in protein solutions prior to trypsin digestion via 10 min of ZnCl and methanol-induced protein precipitation at room temperature (RT). ZASP can remove trypsin digestion and LC-MS incompatible detergents such as SDS, Triton X-100, and urea at high concentrations in solution and unbiasedly recover proteins independent of the amount of protein input. We demonstrated the sensitivity and reproducibility of ZASP in an analysis of samples with 1 μg to 1000 μg of proteins. Compared to commonly used sample preparation methods such as SDC-based in-solution digestion, acetone precipitation, FASP, and SP3, ZASP has proven to be an efficient approach. Here, we present ZASP, a practical, robust, and cost-effective proteomic sample preparation method that can be applied to profile different types of samples.
为了确保下游液相色谱-质谱（LC-MS）分析的最高灵敏度，需要一种通用的蛋白质组学分析样品制备方法，该方法能够实现蛋白质的无偏处理、灵活使用试剂以及低蛋白损失。为了满足这些需求，我们开发了一种 ZnCl 沉淀辅助的样品制备方法（ZASP），该方法可在胰蛋白酶消化之前通过 10 分钟的 ZnCl 和甲醇诱导的室温（RT）下的蛋白质沉淀来去除蛋白质溶液中的苛刻洗涤剂和杂质。ZASP 可以去除高浓度溶液中胰蛋白酶消化和 LC-MS 不兼容的洗涤剂，如 SDS、Triton X-100 和尿素，并能无偏地回收蛋白质，而与蛋白质输入量无关。我们在对 1μg 至 1000μg 蛋白质的样品进行分析中证明了 ZASP 的灵敏度和重现性。与常用的样品制备方法（如基于 SDC 的溶液内消化、丙酮沉淀、FASP 和 SP3）相比，ZASP 被证明是一种有效的方法。在这里，我们提出了 ZASP，这是一种实用、稳健且具有成本效益的蛋白质组学样品制备方法，可应用于不同类型的样品分析。
