In the analysis of ~100 individual cells from three different cell lines, we demonstrate that the N2 chip-based scProteomics platform can robustly quantify ~1500 proteins and reveal membrane protein markers. The tandem mass tag (TMT) pooling step is simplified by adding a microliter droplet on the nested nanowells to combine labeled single-cell samples. The N2 chip reduces reaction volume to 240 single cells on a single microchip.
Herein we describe a nested nanoPOTS (N2) chip to improve protein recovery, operation robustness, and processing throughput for isobaric-labeling-based scProteomics workflow. However, single-cell proteomics is still immature and confronts many technical challenges. Global quantification of protein abundances in single cells could provide direct information on cellular phenotypes and complement transcriptomics measurements.
Nature Communications volume 12, Article number: 6246 ( 2021) High-throughput and high-efficiency sample preparation for single-cell proteomics using a nested nanowell chip
0 kommentar(er)
