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The Consortium for Functional Glycomics’ printed glycan microarray format has replaced the earlier ELISA-based microplate array for routine screening of glycan-binding protein (GBP) specificity. A growing library of natural and synthetic glycans with amino linkers is printed onto NHS-activated glass microscope slides (SCHOTT Nexterion) to form covalent amide linkages in a printed glycan array. The current array (version 3.2) has 406 glycan targets. The Protein-Glycan Interaction Core (H) analyzes investigator-generated lectins, antibodies, suspected GBPs, or microorganisms on the glycan microarray. Initial screening on the glycan microarray is performed with 70 microliters of a 200 microgram/mL solution of GBP. The protocols used for these analyses require flourescent reagents for detecting primary binding to the glycans on the array.
The Protein-Glycan Interaction Core (H) has the ELISA-based plate array which is comprised of a library of biotinylated synthetic and natural ligands attached by a spacer arm to streptavidin-coated microtiter plates. Each unique ligand occupies a set of defined addresses on the array in replicate wells (see plate array version 3.8). The Protein-Glycan Interaction Core (H) at its discretion analyzes some GBPs on the plate array using a screening protocol. The plate array of about 230 glycan targets requires approximately 1 milligram of GBP sample compared to approximately 14 micrograms with the printed array.