Cross-platform comparison of glycan microarray formats

Linlin Wang, Richard D. Cummings, David F. Smith, Margaret Huflejt, Christopher T. Campbell, Jeffrey C. Gildersleeve, Jared Q. Gerlach, Michelle Kilcoyne, Lokesh Joshi, Sonia Serna, Niels Christian Reichardt, Núria Parera Pera, Roland J. Pieters, William Eng, Lara K. Mahal

Research output: Contribution to journalArticle

Abstract

Carbohydrates participate in almost every aspect of biology from protein sorting to modulating cell differentiation and cell-cell interactions. To date, the majority of data gathered on glycan expression has been obtained via analysis with either anti-glycan antibodies or lectins. A detailed understanding of the specificities of these reagents is critical to the analysis of carbohydrates in biological systems. Glycan microarrays are increasingly used to determine the binding specificity of glycan-binding proteins (GBPs). In this study, six different glycan microarray platforms with different modes of glycan presentation were compared using five well-known lectins; concanavalin A, Helix pomatia agglutinin, Maackia amurensis lectin I, Sambucus nigra agglutinin and wheat germ agglutinin. A new method (universal threshold) was developed to facilitate systematic comparisons across distinct array platforms. The strongest binders of each lectin were identified using the universal threshold across all platforms while identification of weaker binders was influenced by platform-specific factors including presentation of determinants, array composition and self-reported thresholding methods. This work compiles a rich dataset for comparative analysis of glycan array platforms and has important implications for the implementation of microarrays in the characterization of GBPs.

Original languageEnglish (US)
Pages (from-to)507-517
Number of pages11
JournalGlycobiology
Volume24
Issue number6
DOIs
StatePublished - Jun 2014

Keywords

  • comparison
  • glycan microarray
  • lectin
  • presentation

ASJC Scopus subject areas

  • Biochemistry

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    Wang, L., Cummings, R. D., Smith, D. F., Huflejt, M., Campbell, C. T., Gildersleeve, J. C., Gerlach, J. Q., Kilcoyne, M., Joshi, L., Serna, S., Reichardt, N. C., Pera, N. P., Pieters, R. J., Eng, W., & Mahal, L. K. (2014). Cross-platform comparison of glycan microarray formats. Glycobiology, 24(6), 507-517. https://doi.org/10.1093/glycob/cwu019