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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lequin R (2005) Enzyme immunoassay (EIA)/enzyme-linked immunosorbent assay (ELISA). Clin Chem 51:2415–2418
Thway T, Salimi-Moosavi H (2014) Evaluating the impact of matrix effects on biomarker assay sensitivity. Bioanalysis 6:1081–1091
Kraly J, Fazal MA, Schoenherr RM et al (2006) Bioanalytical applications of capillary electrophoresis. Anal Chem 78:4097–4110
van den Broek I, van Dongen WD (2015) LC–MS-based quantification of intact proteins: perspective for clinical and bioanalytical applications. Bioanalysis 7:1943–1958
Kočová Vlčková H, Pilařová V, Svobodová P et al (2018) Current state of bioanalytical chromatography in clinical analysis. Analyst 143:1305–1325
Cross TG, Hornshaw MP (2016) Can LC and LC-MS ever replace immunoassays? J Appl Bioanal 2:108–116
Taylor G (1953) Dispersion of soluble matter in solvent flowing slowly through a tube. Proc R Soc A Math Phys Eng Sci 219:186–203
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 82:1793–1802
Cottet H, Biron J-P, Martin M (2007) Taylor dispersion analysis of mixtures. Anal Chem 79:9066–9073
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 89:13487–13493
Ye F, Jensen H, Larsen SW et al (2012) Measurement of drug diffusivities in pharmaceutical solvents using Taylor dispersion analysis. J Pharm Biomed Anal 61:176–183
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 92:203–210
Jensen H, Østergaard J (2010) Flow induced dispersion analysis quantifies noncovalent interactions in nanoliter samples. J Am Chem Soc 132:4070–4071
Jensen H, Larsen SW, Larsen C et al (2013) Physicochemical profiling of drug candidates using Capillary-based techniques. J Drug Deliv Sci Technol 23:333–345
Poulsen NN, Andersen NZ, Østergaard J et al (2015) Flow induced dispersion analysis rapidly quantifies proteins in human plasma samples. Analyst 140:4365–4369
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 88:9056–9061
Dempsey GT, Vaughan JC, Chen KH et al (2011) Evaluation of fluorophores for optimal performance in localization-based super-resolution imaging. Nat Methods 8:1027–1040
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 40:779–788
Cifuentes A, Canalejas P, Diez-Masa JC (1999) Preparation of linear polyacrylamide-coated capillaries. J Chromatogr A 830:423–438
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 67:278–283
Chamieh J, Cottet H (2012) Comparison of single and double detection points Taylor dispersion analysis for monodisperse and polydisperse samples. J Chromatogr A 1241:123–127
United States Pharmacopeial Convention (2017) General chapter <621> chromatography. In: First supplement to United States Pharmacopeia 40-National Formulary 35. United States Pharmacopeial Convention, Rockville
Acknowledgement
This work was supported by The Danish Council for Independent Research grant 11-106647, and the Danish Market Development Fund grant 2016-10649.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Pedersen, M.E., Østergaard, J., Jensen, H. (2019). Flow-Induced Dispersion Analysis (FIDA) for Protein Quantification and Characterization. In: Phillips, T.M. (eds) Clinical Applications of Capillary Electrophoresis. Methods in Molecular Biology, vol 1972. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9213-3_8
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9213-3_8
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9212-6
Online ISBN: 978-1-4939-9213-3
eBook Packages: Springer Protocols