Nanopore toward Genuine Single-Molecule Sensing: Molecular Ping-Pong Technology

Nanopore sensing is a so-called label-free, single-molecule technology; however, multiple events of different molecules are recorded to obtain statistically robust data, which can limit both efficiency and sample use. To overcome these challenges, nanopore molecular ping-pong technology enables precise single-molecule manipulation, reducing systematic and stochastic errors by repeatedly measuring the same molecule. This review introduces the fundamentals and advancements of ping-pong technology, highlighting a recent breakthrough achieving over 10,000 recaptures of a single dsDNA molecule within minutes. This innovation not only minimizes sample requirements, which is critical for nonamplifiable samples, but also significantly enhances experimental precision. While current applications focus on dsDNA, extending this technology to protein and glycan analysis could transform nanopore research. Just as nanopore technology revolutionized DNA sequencing, it holds the potential to drive the development of nanopore-based protein and glycan sequencers, paving the way for groundbreaking advancements in molecular biology and biomedicine.