Challenging the development of a next-generation proteome sequencer

Abstract

In this review, we discuss high-sensitivity proteomics technologies, particularly focusing on ultra-low-input samples required for single-cell analysis and clinical applications. Even in today’s era, where genome sequencing has been fully realized, detailed proteomic analyses remain indispensable for fully understanding biological phenomena. Here, we introduce a novel pipette-tip-based column, “ChocoTip,” which significantly minimizes peptide losses, and describe the improvements in peptide purification performance achieved through its distinctive material design. Additionally, we detail the “two-step solubilization method,” which significantly enhances peptide solubility in nanoscale hydrophilic interaction chromatography, enabling ultra-sensitive LC-MS/MS analysis. Integrating these technological advancements markedly improves the sensitivity, reproducibility, and comprehensiveness of proteomic analyses, laying a crucial foundation for developing next-generation proteome sequencers.