Flow-Induced Dispersion Analysis (FIDA) for Protein Quantification and Characterization

Part of the Methods in Molecular Biology book series (MIMB, volume 1972)


Flow-Induced Dispersion Analysis (FIDA) enables characterization and quantification of proteins under native conditions. FIDA is based on measuring the change in size of a ligand as it selectively interacts with the target protein. The unbound ligand has a relatively small apparent hydrodynamic radius (size), which increase in the presence of the analyte due to binding to the analyte. The Kd of the interaction may be obtained in a titration experiment and the measurement of the apparent ligand size in an unknown sample forms the basis for determining the analyte concentration. The apparent molecular size is measured by Taylor dispersion analysis (TDA) in fused silica capillary capillaries. FIDA is a “ligand-binding” assay and has therefore certain features in common with Enzyme-Linked Immunosorbent Assay (ELISA), Surface Plasmon Resonance (SPR), and Biolayer Interferometry (BLI) based techniques. However, FIDA probes a single in-solution binding event and thus makes assay development straightforward, and the absolute size measurement enables built-in assay quality control. Further, as FIDA does not involve surface chemistries, complications related to nonspecific adsorption of analyte and assay components are minimized enabling direct measurement in, e.g., plasma and serum.

Key words

Auto-antibody detection Binding analysis Dissociation constant Molecular interactions FIDA Flow-induced dispersion analysis Ligand-binding assay Plasma Protein-protein interaction Serum Taylor dispersion analysis 



This work was supported by The Danish Council for Independent Research grant 11-106647, and the Danish Market Development Fund grant 2016-10649.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.FIDA-Tech ApsC/O University of CopenhagenCopenhagenDenmark
  2. 2.Department of PharmacyUniversity of CopenhagenCopenhagenDenmark

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