Determination of Fucose Concentration in a Lectin-Based Displacement Microfluidic Assay
Autor: | Peter Påhlsson, Eva Åström, Nathaniel D. Robinson, Per G. Erlandsson |
---|---|
Rok vydání: | 2019 |
Předmět: |
Competitive binding
Fucose Surface plasmon resonance Protein-carbohydrate interaction Fluorescence detection Microfluidic assay Enzyme-Linked Immunosorbent Assay Bioengineering Applied Microbiology and Biotechnology Biochemistry Article Aleuria aurantia chemistry.chemical_compound Lectins Protein–carbohydrate interactions Binding site Molecular Biology Binding Sites Chromatography biology Chemistry Lactoferrin Biochemistry and Molecular Biology Lectin General Medicine Microfluidic Analytical Techniques biology.organism_classification Fluorescence biology.protein Biokemi och molekylärbiologi Biotechnology |
Zdroj: | Applied Biochemistry and Biotechnology |
ISSN: | 1559-0291 0273-2289 |
DOI: | 10.1007/s12010-018-02944-5 |
Popis: | 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. Funding Agencies|Health Research Council of Southeast of Sweden [FORSS-389021]; LIST grant from Linkoping University |
Databáze: | OpenAIRE |
Externí odkaz: |