Digital molecular kinetics enabled by large-scale nanopore arrays for multidimensional biophysical profiling

DMK using large-scale nanopore arrays recovers SPR association and dissociation rate constants while enabling robust thermodynamic-kinetic profiling across a wide temperature range and resolving heterogeneous conformations.

• Why it matters: SPR is limited by narrow temporal windows, dynamic heterogeneity, and poor physiological temperature performance, while early nanopore methods often suffer from limited throughput and customized modification dependence.
• What they did: The platform uses a universal coupling and probe reporting system to profile diverse molecular pairs (proteins, peptides, small molecules, DNA, and polymerase complexes) with massive single-molecule event analysis.
• The result: DMK enables ultra-low dissociation rate quantification via maximum likelihood estimation, reveals energy wells via voltage gradients, captures rapid binding events missed by SPR, and supports binding analysis under environmental perturbations.