hosphorylation events are key signal transduction mechanisms within the eukaryotic cells and further understanding such signaling
pathways can elucidate crucial molecular signatures of various human diseases (1-3). Te global deep profiling of phosphospeptide has beensuccessfully optimized in the procedures of enrichment, mass spectrometry, and data analysis (4,5). However, phosphopeptide enrichment ishighly labor-intensive and a low-throughput process that leads to poor reproducibility (6-8). Here, we introduce a phosphopeptide enrichmentleveraging dispersive pipette extraction technology on IMCStips. In comparison to conventional solid phase extraction technologies, thepatented dispersive pipette extraction process uses loose resin that mixes during aspiration and dispense steps. Tis increases the interactionbetween the resin and sample via turbulent mixing within the pipette tips. Te tips containing titanium oxide (TiO2) resin were used toexplore the initial feasibility of DPX technology for phosphopeptide enrichment. Te reported method uses automated liquid handlingsystems, such as Integra ViaFlo96, to enrich phosphopeptides from cell lysates for high-throughput and reproducible sample preparation tostudy phosphorylation events.