From Amino Acids to Proteins via Peptide-Centric Nanopore Technology for Sensing, Probing, and Design

Direct, high-resolution analysis of proteins and peptides at the single-molecule level remains a major challenge in biology and medicine. Nanopore technology has emerged as a uniquely powerful tool for label-free, real-time detection of biomolecules. This review comprehensively surveys advances in nanopore sensing, with a focus on peptides as pivotal intermediates between amino acids and full-length proteins. We begin by examining the foundational sensing of individual amino acids, followed by direct peptide analysis encompassing discrimination of isomers, post-translational modifications, and biomarkers. Subsequently, we discuss indirect strategies for protein identification and metal ion detection via peptide probes. Finally, we explore cutting-edge frontiers in peptide sequencing and the de novo design of peptide-based nanopores. With continued development, nanopore sensing holds substantial potential to drive transformative advances in biomolecular analysis, proteomics, and diagnostics.