title: Cross-platform comparison of glycan microarray formats.
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.
authors.affiliation: Biomedical Chemistry Institute, New York University Department of Chemistry, 100 Washington Square East, Room 1001, New York, NY 10003, USA.
authors.forename: Linlin
authors.initials: L
authors.lastname: Wang
authors.forename: Richard D
authors.initials: RD
authors.lastname: Cummings
authors.forename: David F
authors.initials: DF
authors.lastname: Smith
authors.forename: Margaret
authors.initials: M
authors.lastname: Huflejt
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Campbell
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Gildersleeve
authors.forename: Jared Q
authors.initials: JQ
authors.lastname: Gerlach
authors.forename: Michelle
authors.initials: M
authors.lastname: Kilcoyne
authors.forename: Lokesh
authors.initials: L
authors.lastname: Joshi
authors.forename: Sonia
authors.initials: S
authors.lastname: Serna
authors.forename: Niels-Christian
authors.initials: NC
authors.lastname: Reichardt
authors.forename: Núria
authors.initials: N
authors.lastname: Parera Pera
authors.forename: Roland J
authors.initials: RJ
authors.lastname: Pieters
authors.forename: William
authors.initials: W
authors.lastname: Eng
authors.forename: Lara K
authors.initials: LK
authors.lastname: Mahal
mesh_terms: D001665:Binding Sites; D002241:Carbohydrates / Q000096:biosynthesis; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D003208:Concanavalin A / Q000737:chemistry / Q000378:metabolism; D037102:Lectins / Q000737:chemistry / Q000378:metabolism; D046228:Microarray Analysis; D010835:Phytohemagglutinins / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D014909:Wheat Germ Agglutinins / Q000737:chemistry / Q000378:metabolism
chemical_list: D002241:Carbohydrates; D002352:Carrier Proteins; C038095:Helix lectin; D037102:Lectins; D010835:Phytohemagglutinins; D011134:Polysaccharides; D014909:Wheat Germ Agglutinins; C100156:leukoagglutinins, plants; D003208:Concanavalin A
keywords: comparison; glycan microarray; lectin; presentation