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Imaging Analysis of Carbohydrate-Modified Surfaces Using ToF-SIMS and SPRi


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1

Whitman College, 345 Boyer Ave., Walla Walla, WA 99362, USA

2

Department of Bioengineering, University of Washington, Seattle, WA 98195, USA

3

NESAC-Bio, University of Washington, Seattle, WA 98195, USA



These authors contributed equally to this work.





*

Author to whom correspondence should be addressed.



Abstract Covalent modification of surfaces with carbohydrates glycans is a prerequisite for a variety of glycomics-based biomedical applications, including functional biomaterials, glycoarrays, and glycan-based biosensors. The chemistry of glycan immobilization plays an essential role in the bioavailability and function of the surface bound carbohydrate moiety. However, the scarcity of analytical methods to characterize carbohydrate-modified surfaces complicates efforts to optimize glycan surface chemistries for specific applications. Time-of-Flight Secondary Ion Mass Spectrometry ToF-SIMS is a surface sensitive technique suited for probing molecular composition at the biomaterial interface. Expanding ToF-SIMS analysis to interrogate carbohydrate-modified materials would increase our understanding of glycan surface chemistries and advance novel tools in the nascent field of glycomics. In this study, a printed glycan microarray surface was fabricated and subsequently characterized by ToF-SIMS imaging analysis. A multivariate technique based on principal component analysis PCA was used to analyze the ToF-SIMS dataset and reconstruct ToF-SIMS images of functionalized surfaces. These images reveal chemical species related to the immobilized glycan, underlying glycan-reactive chemistries, gold substrates, and outside contaminants. Printed glycoarray elements spots were also interrogated to resolve the spatial distribution and spot homogeneity of immobilized glycan. The bioavailability of the surface-bound glycan was validated using a specific carbohydrate-binding protein lectin as characterized by Surface Plasmon Resonance Imaging SPRi. Our results demonstrate that ToF-SIMS is capable of characterizing chemical features of carbohydrate-modified surfaces and, when complemented with SPRi, can play an enabling role in optimizing glycan microarray fabrication and performance. View Full-Text

Keywords: carbohydrate microarray; ToF-SIMS; SPR; glycomics carbohydrate microarray; ToF-SIMS; SPR; glycomics





Autor: Kathryn M. Bolles 1,†, Fang Cheng 2,†, Jesse Burk-Rafel 2,†, Manish Dubey 3 and Daniel M. Ratner 2,*

Fuente: http://mdpi.com/



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