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Photogenerated carbohydrate microarrays to study carbohydrate-protein interactions using surface plasmon resonance imaging
KTH, Skolan för kemivetenskap (CHE), Kemi, Organisk kemi.
KTH, Skolan för kemivetenskap (CHE), Kemi, Organisk kemi.ORCID-id: 0000-0002-1533-6514
Vise andre og tillknytning
2010 (engelsk)Inngår i: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 26, nr 2, s. 344-350Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

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.

sted, utgiver, år, opplag, sider
2010. Vol. 26, nr 2, s. 344-350
Emneord [en]
Carbohydrate microarray, Photocoupling chemistry, Immobilization, Lectins, Surface plasmon resonance imaging
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-26220DOI: 10.1016/j.bios.2010.08.009ISI: 000283804400008Scopus ID: 2-s2.0-77956904068OAI: oai:DiVA.org:kth-26220DiVA, id: diva2:399936
Forskningsfinansiär
EU, European Research Council, MRTN-CT-19561
Merknad

QC 20150728

Tilgjengelig fra: 2011-02-24 Laget: 2010-11-21 Sist oppdatert: 2019-12-20bibliografisk kontrollert
Inngår i avhandling
1. Photochemical Surface Functionalization: Synthesis, Nanochemistry and Glycobiological Studies
Åpne denne publikasjonen i ny fane eller vindu >>Photochemical Surface Functionalization: Synthesis, Nanochemistry and Glycobiological Studies
2011 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

This thesis mainly deals with the development of photochemical approaches to immobilize carbohydrates on surfaces for glycobiological studies. These approaches have been incorporated into a number of state-of-the-art nanobio-platforms, including carbohydrate microarrays, surface plasmon resonance (SPR), quartz crystal microbalance (QCM), atomic force microscopy (AFM), and glyconanomaterials. All the surfaces have displayed good binding capabilities and selectivities after functionalization with carbohydrates, and a range of important data have been obtained concerning surface characteristics and carbohydrate-protein interactions, based on the platforms established. Besides, a variety of non-carbohydrate and carbohydrate-based molecules have been synthesized, during which process the mutarotation of 1-glycosyl thiols and the stereocontrol in 1-S-glycosylation reactions have been thoroughly studied.

sted, utgiver, år, opplag, sider
Stockholm: KTH Royal Institute of Technology, 2011. s. 86
Serie
Trita-CHE-Report, ISSN 1654-1081 ; 2011:47
Emneord
Carbohydrates, Glycobiology, Carbohydrate-protein interactions, Perfluorophenylazide (PFPA), Photochemistry, Carbohydrate surface immobilization, Carbohydrate microarrays, Surface plasmon resonance (SPR), Quartz crystal microbalance (QCM), Atomic force microscopy (AFM), Glyconanomaterials, Concanavalin A (Con A), Cyanovirin-N (CV-N), 1-Glycosyl thiols; Mutarotation, 1-S-glycosylation, Stereocontrol
HSV kategori
Identifikatorer
urn:nbn:se:kth:diva-41973 (URN)978-91-7501-072-4 (ISBN)
Disputas
2011-10-21, F3, Lindstedtsvägen 26, KTH, Stockholm, 13:00 (engelsk)
Opponent
Veileder
Merknad
QC 20111004Tilgjengelig fra: 2011-10-04 Laget: 2011-10-04 Sist oppdatert: 2011-10-04bibliografisk kontrollert

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