Department of Pharmacology, School of Medicine, CHA University, Seongnam-si, Gyeonggi-do, South Korea.
Proteomics Research Team, CHA Future Medicine Research Institute, Seongnam-si, Gyeonggi-do, South Korea.
Methods Mol Biol. 2024;2823:141-154. doi: 10.1007/978-1-0716-3922-1_10.
Mass spectrometry-based single-cell proteomics has undergone rapid progress and has become an active research area. However, because of the ultralow amount of proteins in single cells, it is still highly challenging to achieve efficient sample preparation and sensitive LC-MS detection. Here, we provide a detailed protocol for isobaric labeling-based single-cell proteomics relying on a microfluidic droplet-based sample processing technology. The protocol allows for processing both single cells and carrier samples in separate microchips using a commercially available platform (cellenONE) with high sample recovery and high throughput. We also provide an optimized LC-MS method for sensitive and robust data collection.
基于质谱的单细胞蛋白质组学发展迅速，已成为一个活跃的研究领域。然而，由于单细胞中蛋白质的含量极低，因此仍然非常难以实现高效的样品制备和灵敏的 LC-MS 检测。在这里，我们提供了一种基于等压标记的单细胞蛋白质组学详细方案，该方案依赖于基于微流控液滴的样品处理技术。该方案允许使用市售平台（cellenONE）在单独的微芯片中处理单个细胞和载体样品，具有高样品回收率和高通量。我们还提供了一种优化的 LC-MS 方法，用于灵敏和稳健的数据采集。
