Abstract
The antibody–lectin sandwich arrays (ALSA) is a powerful new tool for glycoproteomics research. ALSA enables precise measurements of the glycosylation states of multiple proteins captured directly from biological samples. The platform can be used in a high-throughput mode with low sample consumption, making it well suited to biomarker research exploring glycan alterations on specific proteins. This article provides detailed descriptions of the use of ALSA, with a particular focus on biomarker research. The preparation and selection of antibodies and lectins, the preparation and use of the arrays and samples, and special considerations for using the platform for biomarker research are covered.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Chen, S., LaRoche, T., Hamelinck, D., Bergsma, D., Brenner, D., Simeone, D., Brand, R. E., and Haab, B. B. (2007) Multiplexed analysis of glycan variation on native proteins captured by antibody microarrays. Nature methods 4, 437–444.
Yue, T., Goldstein, I. J., Hollingsworth, M. A., Kaul, K., Brand, R. E., and Haab, B. B. (2009) The prevalence and nature of glycan alterations on specific proteins in pancreatic cancer patients revealed using antibody-lectin sandwich arrays. Mol Cell Proteomics 8, 1697–1707.
Dube, D. H., and Bertozzi, C. R. (2005) Glycans in cancer and inflammation--potential for therapeutics and diagnostics. Nat Rev Drug Discov 4, 477–488.
Hirabayashi, J. (2004) Lectin-based structural glycomics: glycoproteomics and glycan profiling. Glycoconjugate journal 21, 35–40.
Yue, T., and Haab, B. B. (2009) Microarrays in glycoproteomics research. Clin Lab Med 29, 15–29.
Haab, B. B., and Zhou, H. (2004) Multiplexed protein analysis using spotted antibody microarrays. Methods in molecular biology Clifton, N.J 264, 33–45.
Chen, S., and Haab, B. B. (2007) Antibody Microarrays for Protein and Glycan Detection. In: Van Eyk, J., and Dunn, M., eds. Clinical Proteomics, Wiley, VCH, Weinheim, Germany.
Haab, B. B. (2005) Multiplexed protein analysis using antibody microarrays and label-based detection. Methods Mol Med 114, 183–194.
Haab, B. B., and Lizardi, P. M. (2006) RCA-enhanced protein detection arrays. Methods in molecular biology Clifton, N.J 328, 15–29.
Hirabayashi, J. (2008) Concept, strategy and realization of lectin-based glycan profiling. Journal of biochemistry 144, 139–147.
Culf, A. S., Cuperlovic-Culf, M., and Ouel-lette, R. J. (2006) Carbohydrate microarrays: survey of fabrication techniques. Omics 10, 289–310.
Wang, D. (2003) Carbohydrate microarrays. Proteomics 3, 2167–2175.
Blixt, O., Head, S., Mondala, T., Scanlan, C., Huflejt, M. E., Alvarez, R., Bryan, M. C., Fazio, F., Calarese, D., Stevens, J., Razi, N., Stevens, D. J., Skehel, J. J., van Die, I., Burton, D. R., Wilson, I. A., Cummings, R., Bovin, N., Wong, C. H., and Paulson, J. C. (2004) Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proceedings of the National Academy of Sciences of the United States of America 101, 17033–17038.
Porter, A., Yue, T., Heeringa, L., Day, S., Suh, E., and Haab, B. B. (2009) A motif-based analysis of glycan array data to determine the specificities of glycan-binding proteins. Glycobiology.
Chen, S., and Haab, B. B. (2009) Analysis of glycans on serum proteins using antibody microarrays. Methods in molecular biology Clifton, N.J 520, 39–58.
Rai, A. J., Gelfand, C. A., Haywood, B. C., Warunek, D. J., Yi, J., Schuchard, M. D., Mehigh, R. J., Cockrill, S. L., Scott, G. B., Tammen, H., Schulz-Knappe, P., Speicher, D. W., Vitzthum, F., Haab, B. B., Siest, G., and Chan, D. W. (2005) HUPO Plasma Proteome Project specimen collection and handling: towards the standardization of parameters for plasma proteome samples. Proteomics 5, 3262–3277.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Haab, B.B., Yue, T. (2011). High-Throughput Studies of Protein Glycoforms Using Antibody–Lectin Sandwich Arrays. In: Korf, U. (eds) Protein Microarrays. Methods in Molecular Biology, vol 785. Humana Press. https://doi.org/10.1007/978-1-61779-286-1_15
Download citation
DOI: https://doi.org/10.1007/978-1-61779-286-1_15
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-285-4
Online ISBN: 978-1-61779-286-1
eBook Packages: Springer Protocols