Abstract
Monitoring multiple biological interactions in a multiplexed array format has numerous advantages. However, converting well-developed surface chemistry for spectroscopic measurements to array-based, high-throughput screening is not a trivial process and often proves to be the bottleneck in method development. This chapter reports the fabrication and characterization of a new carbohydrate microarray with synthetic sialosides for surface plasmon resonance imaging analysis of lectin–carbohydrate interactions. Contact printing of functional sialosides on neutravidin-coated surfaces was carried out and the properties of the resulting elements were characterized by fluorescence microscopy. Sambucus nigra agglutinin (SNA) was used for testing on four different carbohydrate-functionalized surfaces and differential binding was analyzed. Multiplexed detection of SNA/biotinylated sialoside interactions on arrays up to 400 elements has been performed with good data correlation, demonstrating the effectiveness of the biotin–neutravidin-based biointerface to control probe orientation for reproducible and efficient protein binding to carbohydrates.
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Acknowledgments
The authors would like to acknowledge the financial supports from NSF grant CHE-0719224 (to QC) and NIH grant R01GM076360 (to XC). MJL would like to acknowledge the support from the American Chemical Society, Division of Analytical Chemistry Fellowship, sponsored by Procter & Gamble.
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Linman, M.J., Yu, H., Chen, X., Cheng, Q. (2012). Surface Plasmon Resonance Imaging Analysis of Protein Binding to a Sialoside-Based Carbohydrate Microarray. In: Chevolot, Y. (eds) Carbohydrate Microarrays. Methods in Molecular Biology, vol 808. Humana Press. https://doi.org/10.1007/978-1-61779-373-8_13
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DOI: https://doi.org/10.1007/978-1-61779-373-8_13
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