Product Isolation

For hydrophobic products like glycolipids or other aliphatic aglycones, C18 reversed phase supports can be used for their isolation from enzyme reactions. In milligram-scale synthesis, C18 Sep-Pak cartridges from Waters perform well. The resin or cartridge should be preconditioned before use by washing with MeOH, then equilibrated with water before aqueous enzymatic reaction samples are loaded. Organic solvents such as glycerol or DMSO in enzymatic reactions prevent substrates from binding to the C18 resin. In these cases, a dilution with water to less than 1% organic solvent in the enzymatic samples prior to loading on columns is recommended.

Ion exchange chromatography can be used for the isolation of sialylated products and oligosaccharides with amine groups. Size-exclusion chromatography serves well for the isolation of reducing sugars or glycoconjugates with high molecular weights. Adsorption on charcoal is another practical method for the purification of oligosaccharides. A recent publication focuses on the separation of oligosaccharides [132].

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