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September 10, 2025

FIDA-Based Screening of De Novo Binders in Bacterial Lysates and Purified Protein Titration v1

Key Points

FIDA identifies high-affinity biomolecular binders rapidly in complex biological samples like cell lysates, enhancing screening efficiency.
By measuring the hydrodynamic radius of fluorescently labeled molecules, FIDA provides direct detection of binding events in a single measurement.
The low sample requirements of FIDA make it compatible with crude preparations, enabling evaluation of extensive designs without immobilization.
This method allows for quantitative affinity determination through titration following initial screening of candidates.

Abstract

Flow-induced dispersion analysis (FIDA) offers a rapid and cost-effective approach for screening and characterizing biomolecular binders. By measuring changes in the hydrodynamic radius of fluorescently labelled molecules and their complexes, FIDA enables direct detection of binding events in complex biological samples, such as heat-treated cell lysates, without immobilization. This allows identification of candidates with high affinity and stability in a single measurement, followed by quantitative affinity determination through titration. The low sample requirements and compatibility with crude preparations make FIDA a versatile tool for evaluating large sets of computational or experimentally generated binder designs.

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