Flow-induced dispersion analysis
Flow-induced dispersion analysis (FIDA) is an immobilization-free technology used for characterization and quantification of biomolecular interaction and protein concentration under native conditions[1][2][3]. In the FIDA assay, the size of a ligand (indicator) with affinity to the target analyte is measured. When the indicator interacts with the analyte the apparent size increases and this change in size can be used to determine the analyte concentration and interaction[1][2][4]. Additionally, the hydrodynamic radius of the analyte-indicator complex is obtained. A FIDA assay is typically completed in minutes and only requires a modest sample consumption of a few µl[1].
Applications
- Quantification of analytes (e.g. proteins, peptides and DNA) in complex solutions (e.g. plasma and fermentation broth [5][6])
- Determination of
- affinity constants
- binding kinetics
- molecular size (hydrodynamic radius)
- diffusion coefficient
Principle
The FIDA principle is based on measuring the change in the apparent size (diffusivity) of a selective indicator interacting with the analyte molecule[1][2][4]. The apparent indicator size is measured by Taylor dispersion analysis in a capillary under hydrodynamic flow[7].
References
- ^ a b c d e Poulsen, Nicklas N.; Andersen, Nina Z.; Østergaard, Jesper; Zhuang, Guisheng; Petersen, Nickolaj J.; Jensen, Henrik (2015-06-15). "Flow induced dispersion analysis rapidly quantifies proteins in human plasma samples". The Analyst. 140 (13): 4365–4369. doi:10.1039/c5an00697j. ISSN 1364-5528. PMID 26031223.
- ^ a b c Morten E, Pedersen; Østergaard, Jesper; Jensen, Henrik (2019). "Flow-Induced Dispersion Analysis (FIDA) for Protein Quantification and Characterization". In Phillips, Terry M. (ed.). Clinical Applications of Capillary Electrophoresis: Methods and Protocols. Methods in Molecular Biology. Vol. 1972. New York, NY: Springer New York. pp. 109–123. doi:10.1007/978-1-4939-9213-3. ISBN 9781493992126.
- ^ Pedersen, Morten E.; Gad, Sarah I.; Østergaard, Jesper; Jensen, Henrik (2019-04-05). "Protein Characterization in 3D: Size, Folding, and Functional Assessment in a Unified Approach". Analytical Chemistry. 91 (8): 4975–4979. doi:10.1021/acs.analchem.9b00537. ISSN 0003-2700. PMID 30916933.
- ^ a b Jensen, Henrik; Østergaard, Jesper (2010-03-31). "Flow Induced Dispersion Analysis Quantifies Noncovalent Interactions in Nanoliter Samples". Journal of the American Chemical Society. 132 (12): 4070–4071. doi:10.1021/ja100484d. ISSN 0002-7863. PMID 20201527.
- ^ Poulsen, Nicklas N.; Pedersen, Morten E.; Østergaard, Jesper; Petersen, Nickolaj J.; Nielsen, Christoffer T.; Heegaard, Niels H. H.; Jensen, Henrik (2016-09-20). "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". Analytical Chemistry. 88 (18): 9056–9061. doi:10.1021/acs.analchem.6b01741. ISSN 0003-2700. PMID 27571264.
- ^ Pedersen, Morten E.; Østergaard, Jesper; Jensen, Henrik (2020-04-28). "In-Solution IgG Titer Determination in Fermentation Broth Using Affibodies and Flow-Induced Dispersion Analysis". ACS Omega. doi:10.1021/acsomega.0c00791. ISSN 2470-1343.
- ^ Taylor, Sir Geoffrey; S, F. R. (1953-08-25). "Dispersion of soluble matter in solvent flowing slowly through a tube". Proc. R. Soc. Lond. A. 219 (1137): 186–203. doi:10.1098/rspa.1953.0139. ISSN 0080-4630.