Array-Based Functional Screening of Heparin Glycans

SummaryArray methodologies have become powerful tools for interrogation of glycan-protein interactions but have critically lacked the ability to generate cell response data. Here, we report the development of a slide-based array method exemplified by measurement of activation of fibroblast growth factor signaling by heparin saccharides. Heparan sulfate-deficient Swiss 3T3 cells were overlaid onto an aminosilane-coated slide surface onto which heparin saccharides had been spotted and immobilized. The cells were transiently stimulated with FGF2 and immunofluorescence measured to assess downstream ERK1/2 phosphorylation. Activation of this signaling pathway response was restricted to cells exposed to heparin saccharides competent to activate FGF2 signaling. Differential activation of the overlaid cells by different-sized heparin saccharides was demonstrated by quantitative measurement of fluorescence intensity. This “glycobioarray” platform has significant potential as a generic tool for functional glycomics screening.

Graphical AbstractFigure optionsDownload full-size imageDownload high-quality image (160 K)Download as PowerPoint slideHighlights► Current glycoarrays generate data on interactions but not cell responses ► A slide-based method was developed for measuring responses of cells to glycans ► Signaling responses to immobilized glycans can be measured by immunofluorescence ► This glycobioarray platform has generic potential as a tool for functional glycomics