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2014 ; 24
(6
): 507-17
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Cross-platform comparison of glycan microarray formats
#MMPMID24658466
Wang L
; Cummings RD
; Smith DF
; Huflejt M
; Campbell CT
; Gildersleeve JC
; Gerlach JQ
; Kilcoyne M
; Joshi L
; Serna S
; Reichardt NC
; Parera Pera N
; Pieters RJ
; Eng W
; Mahal LK
Glycobiology
2014[Jun]; 24
(6
): 507-17
PMID24658466
show ga
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.
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