Chemistry of natural glycan microarrays

• Natural glycan microarray is a crucial technology for protein/glycan interactions.
• Expanding natural glycan libraries is the most important task for the field.
• Chemo-enzymatic synthesis applied to natural glycans will expand glycan array diversity.
• Presentation of glycans could affect sensitivity of detection of GBP binding.

Glycan microarrays have become indispensable tools for studying protein–glycan interactions. Along with chemo-enzymatic synthesis, glycans isolated from natural sources have played important roles in array development and will continue to be a major source of glycans. N-glycans and O-glycans from glycoproteins, and glycans from glycosphingolipids (GSLs) can be released from corresponding glycoconjugates with relatively mature methods, although isolation of large numbers and quantities of glycans is still very challenging. Glycosylphosphatidylinositol (GPI) anchors and glycosaminoglycans (GAGs) are less represented on current glycan microarrays. Glycan microarray development has been greatly facilitated by bifunctional fluorescent linkers, which can be applied in a ‘Shotgun Glycomics’ approach to incorporate isolated natural glycans. Glycan presentation on microarrays may affect glycan binding by GBPs, often through multivalent recognition by the GBP.