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Determination of Fucose Concentration in a Lectin-Based Displacement Microfluidic Assay
Linköping University, Department of Physics, Chemistry and Biology, Surface Physics and Chemistry. Linköping University, Faculty of Science & Engineering.
Linköping University, Department of Clinical and Experimental Medicine, Division of Cell Biology. Linköping University, Faculty of Medicine and Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine, Division of Cell Biology. Linköping University, Faculty of Medicine and Health Sciences.
Linköping University, Department of Physics, Chemistry and Biology, Sensor and Actuator Systems. Linköping University, Faculty of Science & Engineering.ORCID iD: 0000-0002-2773-5096
2019 (English)In: Applied Biochemistry and Biotechnology, ISSN 0273-2289, E-ISSN 1559-0291, Vol. 188, no 3, p. 868-877Article in journal (Refereed) Published
Abstract [en]

We compare three different methods to quantify the monosaccharide fucose in solutions using the displacement of a large glycoprotein, lactoferrin. Two microfluidic analysis methods, namely fluorescence detection of (labeled) lactoferrin as it is displaced by unlabeled fucose and the displacement of (unlabeled) lactoferrin in SPR, provide fast responses and continuous data during the experiment, theoretically providing significant information regarding the interaction kinetics between the saccharide groups and binding sites. For comparison, we also performed a static displacement ELISA. The stationary binding site in all cases was immobilized S2-AAL, a monovalent polypeptide based on Aleuria aurantia lectin. Although all three assays showed a similar dynamic range, the microfluidic assays with fluorescent or SPR detection show an advantage in short analysis times. Furthermore, the microfluidic displacement assays provide a possibility to develop a one-step analytical platform.

Place, publisher, year, edition, pages
SPRINGER , 2019. Vol. 188, no 3, p. 868-877
Keywords [en]
Competitive binding; Fucose; Surface plasmon resonance; Protein-carbohydrate interaction; Fluorescence detection; Microfluidic assay
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:liu:diva-158829DOI: 10.1007/s12010-018-02944-5ISI: 000471851400019PubMedID: 30710193OAI: oai:DiVA.org:liu-158829DiVA, id: diva2:1337684
Note

Funding Agencies|Health Research Council of Southeast of Sweden [FORSS-389021]; LIST grant from Linkoping University

Available from: 2019-07-16 Created: 2019-07-16 Last updated: 2019-07-16

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