Abstract
Accurate characterization of molecular interactions is fundamental to understanding biological systems and developing therapeutic strategies. Conventional surface-based methods, however, often introduce immobilization artifacts, distorting native binding behavior, reducing flexibility, and making assay development time-consuming.
Here, we present Microfluidic Diffusional Sizing (MDS), an innovative analytical technique that measures protein interactions directly in solution. By avoiding immobilization, MDS enables advanced assays that can simultaneously determine both affinity and calibration-free analyte concentration. These assays benefit from integrated Bayesian analysis, which provides a complete view of parameter space and streamlines experimental design for accurate results with minimal resources.