Photochemical Ligation Techniques for Carbohydrate Biosensors and Protein Interaction Studies
2012 (English)Doctoral thesis, comprehensive summary (Other academic)
This thesis concerns the development of surface ligation techniques for the preparation of carbohydrate biosensors. Several methodologies were developed based on efficient photochemical insertion reactions which quickly functionalize polymeric materials, with either carbohydrates or functional groups such as alkynes or alkenes. The alkyne/alkene surfaces were then treated with carbohydrate azides or thiols and reacted under chemoselective Cu-catalyzed azide-alkyne cycloaddition (CuAAC) or photo-radical thiol-ene/yne click chemistry, thus creating a range of carbohydrate biosensor surfaces under ambient conditions. The methodologies were evaluated by quartz crystal microbalance (QCM) and surface plasmon resonance (SPR) flow through instrumentations with recurring injections of a range of lectins, allowing for real-time analysis of the surface interactions. The developed methods were proven robust and versatile, and the generated carbohydrate biosensors showed high specificities and good capacities for lectin binding. The methods were then used to investigate how varying the glycan linker length and/or a sulfur-linkage affect the subsequent protein binding. The survey was further explored by investigating the impact of sulfur in glycosidic linkages on protein binding, through competition assays with various O/S-linked disaccharides in solution interactions with lectins.
Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2012. , xi, 89 p.
Trita-CHE-Report, ISSN 1654-1081 ; 2012:10
Carbohydrates, Biosensors, CuAAC, Photochemistry, Thiol-ene/yne, Perfluorophenylazide (PFPA), Lectins, Molecular recognition, Quartz crystal microbalance (QCM), Surface plasmon resonance (SPR)
IdentifiersURN: urn:nbn:se:kth:diva-90956ISBN: 978-91-7501-271-1OAI: oai:DiVA.org:kth-90956DiVA: diva2:508185
2012-03-30, F3, Lindstedtsvägen 26, Stockholm, 10:00 (English)
Vincent, Stéphane, Prof.
Ramström, Olof, Professor
QC 201203092012-03-092012-03-052012-03-09Bibliographically approved
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