The SpyBLI cell-free pipeline for the rapid quantification of binding kinetics from crude samples.

ABSTRACT: Accurate measurements of binding kinetics, encompassing equilibrium dissociation constant (K _D), association rate (k _on), and dissociation rate (k _off), are critical for the development and optimisation of high-affinity binding proteins. However, such measurements require highly purified material and precise ligand immobilisation, limiting the number of binders that can be characterised within a reasonable timescale and budget. Here, we present the SpyBLI method, a rapid and cost-effective biolayer interferometry (BLI) pipeline that leverages the SpyCatcher003-SpyTag003 covalent association, eliminating the need for both binder purification and concentration determination. This approach allows for accurate binding-kinetic measurements t

SUBMITTER: Predeina O 

PROVIDER: S-EPMC12247212 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature