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Fabrication of carbohydrate surfaces by using non-derivatised oligosaccharides

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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. © 2012 Springer Science+Business Media, LLC.

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Popplewell, J., Swann, M., Brown, G., & Lauder, B. (2012). Fabrication of carbohydrate surfaces by using non-derivatised oligosaccharides. Methods in Molecular Biology, 808, 221–229. https://doi.org/10.1007/978-1-61779-373-8_15

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