biostudies-literaturePinheiro FDanmarks GrundforskningsfondDanmarks Frie ForskningsfondLundbeck Foundatione70286https://www.ebi.ac.uk/biostudies/studies/S-EPMC12432402biostudies-literatureUnknown34(10)Computational protein design can create binders against targets of interest, but identifying binders with sufficient affinity still requires biochemical screening of many designs. In this work, we test flow-induced dispersion analysis (FIDA) as a method for screening binders in a time and cost-effective manner. FIDA uses Taylor dispersion analysis to determine the hydrodynamic radius of fluorescently labeled biomolecules and their complexes. Here, we use FIDA to assess the binding of RFdiffusion-designed protein binders against the small helical peptide ALFA-tag and the guanylate kinase (GK) domain of PSD-95. Successful binders can be identified in a single measurement using heat-treated bacterial lysates, allowing rapid identification of binders with high affinity and thermostability. SubProtein science : a publication of the Protein SocietyScreening de novo designed protein binders in unpurified lysate using flow induced dispersion analysis.PMC12432402026-00069BDNRF133R449-2023-1396Nowak JSPheasant ECZueva ELampinen VKjaergaard MPinheiro FGrene IKfalseScreening de novo designed protein binders in unpurified lysate using flow induced dispersion analysis.Computational protein design can create binders against targets of interest, but identifying binders with sufficient affinity still requires biochemical screening of many designs. In this work, we test flow-induced dispersion analysis (FIDA) as a method for screening binders in a time and cost-effective manner. FIDA uses Taylor dispersion analysis to determine the hydrodynamic radius of fluorescently labeled biomolecules and their complexes. Here, we use FIDA to assess the binding of RFdiffusion-designed protein binders against the small helical peptide ALFA-tag and the guanylate kinase (GK) domain of PSD-95. Successful binders can be identified in a single measurement using heat-treated bacterial lysates, allowing rapid identification of binders with high affinity and thermostability. Sub2025-01-01T00:00:00Z2025 Oct2026-06-03T03:03:25.659Z2026-04-23T03:13:14.685ZS-EPMC124324024094442210.1002/pro.70286