Photogenerated carbohydrate microarrays to study carbohydrate-protein interactions using surface plasmon resonance imaging
2010 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 26, no 2, 344-350 p.Article in journal (Refereed) Published
A photochemical strategy to generate carbohydrate microarrays on flat sensor surfaces, and to study the protein-binding effects of these arrays by surface plasmon resonance imaging is described. The approach was validated using a panel of carbohydrate-binding proteins. The coupling agents, thiol-functionalized perfluorophenyl azides, allow the covalent attachment of underivatized carbohydrates to gold surfaces by a fast photochemical reaction. Carbohydrate microarrays composed of 3,6-di-O-(alpha-D-mannopyranosyl)-D-mannopyranose (Man3), 2-O-alpha-D-mannopyranosyl-D-mannopyranose (Man2), D-mannose (Man), D-glucose (Glc), and D-galactose (Gal) were constructed, and the binding studies were carried out in real-time using surface plasmon resonance imaging. Results showed that the immobilized carbohydrate ligands retained their binding affinities with lectins, the rank order of which was consistent with that of the free ligands in solution. The detection limit of Man3, Man2, Man, and Glc with the lectin Concanavalin A was measured to be 0.29 nM, 0.18 nM, 0.61 nM, and 3.1 nM, respectively. In addition, soybean agglutinin and Griffonia simplicifolia lectin II were tested on the array, and the results were consistent with the binding selectivity of these lectins with the carbohydrate ligands.
Place, publisher, year, edition, pages
2010. Vol. 26, no 2, 344-350 p.
Carbohydrate microarray, Photocoupling chemistry, Immobilization, Lectins, Surface plasmon resonance imaging
IdentifiersURN: urn:nbn:se:kth:diva-26220DOI: 10.1016/j.bios.2010.08.009ISI: 000283804400008ScopusID: 2-s2.0-77956904068OAI: oai:DiVA.org:kth-26220DiVA: diva2:399936
FunderEU, European Research Council, MRTN-CT-19561
QC 201507282011-02-242010-11-212015-07-28Bibliographically approved