An automated platform for the enzyme-mediated assembly of complex oligosaccharides

Nat Chem. 2019 Mar;11(3):229-236. doi: 10.1038/s41557-019-0219-8. Epub 2019 Feb 21.

Abstract

An automated platform that can synthesize a wide range of complex carbohydrates will greatly increase their accessibility and should facilitate progress in glycoscience. Here we report a fully automated process for enzyme-mediated oligosaccharide synthesis that can give easy access to different classes of complex glycans including poly-N-acetyllactosamine derivatives, human milk oligosaccharides, gangliosides and N-glycans. Our automated platform uses a catch and release approach in which glycosyltransferase-catalysed reactions are performed in solution and product purification is accomplished by solid phase extraction. We developed a sulfonate tag that can easily be installed and enables highly efficient solid phase extraction and product release using a single set of washing conditions, regardless of the complexity of the glycan. Using this custom-built synthesizer, as many as 15 reaction cycles can be performed in an automated fashion without a need for lyophilization or buffer exchange steps.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Carbohydrate Sequence
  • Chemistry Techniques, Synthetic / methods*
  • Gangliosides
  • Glycosyltransferases / metabolism*
  • Humans
  • Milk, Human
  • Oligosaccharides* / chemical synthesis
  • Oligosaccharides* / chemistry
  • Oligosaccharides* / metabolism
  • Polysaccharides
  • Solid Phase Extraction

Substances

  • Gangliosides
  • Oligosaccharides
  • Polysaccharides
  • poly-N-acetyllactosamine
  • Glycosyltransferases