Advertisement

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

  • Morten E. Pedersen
  • Jesper Østergaard
  • Henrik JensenEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1972)

Abstract

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 

Notes

Acknowledgement

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

References

  1. 1.
    Lequin R (2005) Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin Chem 51:2415–2418CrossRefGoogle Scholar
  2. 2.
    Thway T, Salimi-Moosavi H (2014) Evaluating the impact of matrix effects on biomarker assay sensitivity. Bioanalysis 6:1081–1091CrossRefGoogle Scholar
  3. 3.
    Kraly J, Fazal MA, Schoenherr RM et al (2006) Bioanalytical applications of capillary electrophoresis. Anal Chem 78:4097–4110CrossRefGoogle Scholar
  4. 4.
    van den Broek I, van Dongen WD (2015) LC–MS-based quantification of intact proteins: perspective for clinical and bioanalytical applications. Bioanalysis 7:1943–1958CrossRefGoogle Scholar
  5. 5.
    Kočová Vlčková H, Pilařová V, Svobodová P et al (2018) Current state of bioanalytical chromatography in clinical analysis. Analyst 143:1305–1325CrossRefGoogle Scholar
  6. 6.
    Cross TG, Hornshaw MP (2016) Can LC and LC-MS ever replace immunoassays? J Appl Bioanal 2:108–116CrossRefGoogle Scholar
  7. 7.
    Taylor G (1953) Dispersion of soluble matter in solvent flowing slowly through a tube. Proc R Soc A Math Phys Eng Sci 219:186–203Google Scholar
  8. 8.
    Cottet H, Biron J-P, Cipelletti L et al (2010) Determination of individual diffusion coefficients in evolving binary mixtures by Taylor dispersion analysis: application to the monitoring of polymer reaction. Anal Chem 82:1793–1802CrossRefGoogle Scholar
  9. 9.
    Cottet H, Biron J-P, Martin M (2007) Taylor dispersion analysis of mixtures. Anal Chem 79:9066–9073CrossRefGoogle Scholar
  10. 10.
    Chamieh J, Leclercq L, Martin M et al (2017) Limits in size of Taylor dispersion analysis: representation of the different hydrodynamic regimes and application to the size-characterization of cubosomes. Anal Chem 89:13487–13493CrossRefGoogle Scholar
  11. 11.
    Ye F, Jensen H, Larsen SW et al (2012) Measurement of drug diffusivities in pharmaceutical solvents using Taylor dispersion analysis. J Pharm Biomed Anal 61:176–183CrossRefGoogle Scholar
  12. 12.
    Jensen SS, Jensen H, Cornett C et al (2014) Insulin diffusion and self-association characterized by real-time UV imaging and Taylor dispersion analysis. J Pharm Biomed Anal 92:203–210CrossRefGoogle Scholar
  13. 13.
    Jensen H, Østergaard J (2010) Flow induced dispersion analysis quantifies noncovalent interactions in nanoliter samples. J Am Chem Soc 132:4070–4071CrossRefGoogle Scholar
  14. 14.
    Jensen H, Larsen SW, Larsen C et al (2013) Physicochemical profiling of drug candidates using Capillary-based techniques. J Drug Deliv Sci Technol 23:333–345CrossRefGoogle Scholar
  15. 15.
    Poulsen NN, Andersen NZ, Østergaard J et al (2015) Flow induced dispersion analysis rapidly quantifies proteins in human plasma samples. Analyst 140:4365–4369CrossRefGoogle Scholar
  16. 16.
    Poulsen NN, Pedersen ME, Østergaard J et al (2016) Flow-induced dispersion analysis for probing anti-dsDNA antibody binding heterogeneity in systemic lupus erythematosus patients: toward a new approach for diagnosis and patient stratification. Anal Chem 88:9056–9061CrossRefGoogle Scholar
  17. 17.
    Dempsey GT, Vaughan JC, Chen KH et al (2011) Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging. Nat Methods 8:1027–1040CrossRefGoogle Scholar
  18. 18.
    Poulsen NN, Østergaard J, Petersen NJ et al (2017) Automated coating procedures to produce poly(ethylene glycol) brushes in fused-silica capillaries. J Sep Sci 40:779–788CrossRefGoogle Scholar
  19. 19.
    Cifuentes A, Canalejas P, Diez-Masa JC (1999) Preparation of linear polyacrylamide-coated capillaries. J Chromatogr A 830:423–438CrossRefGoogle Scholar
  20. 20.
    Baderia VK, Gowri VS, Sanghi SK et al (2012) Stable physically adsorbed coating of poly-vinyl alcohol for the separation of basic proteins. J Anal Chem 67:278–283CrossRefGoogle Scholar
  21. 21.
    Chamieh J, Cottet H (2012) Comparison of single and double detection points Taylor dispersion analysis for monodisperse and polydisperse samples. J Chromatogr A 1241:123–127CrossRefGoogle Scholar
  22. 22.
    United States Pharmacopeial Convention (2017) General chapter <621> chromatography. In: First supplement to United States Pharmacopeia 40-National Formulary 35. United States Pharmacopeial Convention, RockvilleGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Morten E. Pedersen
    • 1
  • Jesper Østergaard
    • 1
    • 2
  • Henrik Jensen
    • 1
    • 2
    Email author
  1. 1.FIDA-Tech ApsC/O University of CopenhagenCopenhagenDenmark
  2. 2.Department of PharmacyUniversity of CopenhagenCopenhagenDenmark

Personalised recommendations