Biology:Flow-induced dispersion analysis
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The FIDA principle: A narrow indicator zone is introduced into a capillary under hydrodynamic flow. When the indicator is not bound, a narrow peak is observed at the detector. However, when the indicator is bound by the target analyte, the apparent size increases and a broader peak is observed. This change in size can be used for determine the analyte concentration and interaction[1] - Published by The Royal Society of Chemistry.
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, DNA, nanoparticles[5]) in complex solutions (e.g. plasma and fermentation broth [6][7])
- Determination of
- affinity constants
- binding kinetics
- molecular size (hydrodynamic radius)
- oligomeric state[8]
- 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.[9]
References
- ↑ 1.0 1.1 1.2 1.3 1.4 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" (in en). The Analyst 140 (13): 4365–4369. doi:10.1039/c5an00697j. ISSN 1364-5528. PMID 26031223.
- ↑ 2.0 2.1 2.2 Morten E, Pedersen; Østergaard, Jesper; Jensen, Henrik (2019). "Flow-Induced Dispersion Analysis (FIDA) for Protein Quantification and Characterization". in Phillips, Terry M.. Clinical Applications of Capillary Electrophoresis: Methods and Protocols. Methods in Molecular Biology. 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" (in en). Analytical Chemistry 91 (8): 4975–4979. doi:10.1021/acs.analchem.9b00537. ISSN 0003-2700. PMID 30916933.
- ↑ 4.0 4.1 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.
- ↑ Cholak, Ersoy; Bugge, Katrine; Khondker, Adree; Gauger, Kimmie; Pedraz‐Cuesta, Elena; Pedersen, Morten Enghave; Bucciarelli, Saskia; Vestergaard, Bente et al. (2020). "Avidity within the N-terminal anchor drives α-synuclein membrane interaction and insertion" (in en). The FASEB Journal 34 (6): 7462–7482. doi:10.1096/fj.202000107R. ISSN 1530-6860. PMID 32277854.
- ↑ 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 5 (18): 10519–10524. doi:10.1021/acsomega.0c00791. ISSN 2470-1343. PMID 32426609.
- ↑ Pedersen, Morten E.; Haegebaert, Ragna M. S.; Østergaard, Jesper; Jensen, Henrik (2021-02-26). "Size-based characterization of adalimumab and TNF-α interactions using flow induced dispersion analysis: assessment of avidity-stabilized multiple bound species" (in en). Scientific Reports 11 (1): 4754. doi:10.1038/s41598-021-84113-z. ISSN 2045-2322. PMID 33637878.
- ↑ Taylor, Sir Geoffrey; S, F. R. (1953-08-25). "Dispersion of soluble matter in solvent flowing slowly through a tube" (in en). Proc. R. Soc. Lond. A 219 (1137): 186–203. doi:10.1098/rspa.1953.0139. ISSN 0080-4630.
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