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Fabrication of carbohydrate surfaces by using non-derivatised oligosaccharides

Popplewell, Jonathan and Swann, Marcus and Brown, Gavin and Lauder, Bob (2012) Fabrication of carbohydrate surfaces by using non-derivatised oligosaccharides. Methods in molecular biology (Clifton, N.J.), 808. pp. 221-229.

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Abstract

Surface-based tools, such as microarrays and optical biosensors, are being increasingly applied to the analysis of carbohydrate-protein interactions. A key to these developments is the presentation of the carbohydrate to the protein target. Dual polarisation interferometry (DPI) is a surface-based technique that permits the real-time measurement of the changes in thickness, refractive index, and mass of adsorbates 100-nm thick or less on the surface of a functionalised waveguide. DPI has been used to design and characterise a surface on which the orientation and density of the immobilised carbohydrates are suitable for studying their interactions with proteins and where non-specific binding is reduced to less than 5% of total binding. A thiol-functionalised surface was derivatised with a heterobifunctional cross-linker to yield a hydrazide surface. This was treated with oligosaccharides, derived from keratan sulphate, chondroitin sulphate, and heparin that possess a reducing end. To block the unreacted hydrazide groups, the surface was treated with an aldehyde-functionalised PEG, and the surfaces were then challenged with a variety of proteins.

Item Type: Article
Journal or Publication Title: Methods in molecular biology (Clifton, N.J.)
Uncontrolled Keywords: Chondroitin ; Keratan
Subjects:
Departments: Faculty of Health and Medicine > Biomedical & Life Sciences
ID Code: 51373
Deposited By: ep_importer_pure
Deposited On: 22 Nov 2011 16:50
Refereed?: Yes
Published?: Published
Last Modified: 09 Apr 2014 22:51
Identification Number:
URI: http://eprints.lancs.ac.uk/id/eprint/51373

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