Unit

Experimental Design Considerations for In Vitro Non-Natural Glycan Display via Metabolic Oligosaccharide Engineering

  1. Elaine Tan,
  2. Ruben T. Almaraz,
  3. Hargun S. Khanna,
  4. Jian Du,
  5. Kevin J. Yarema

Published Online: 1 SEP 2010

DOI: 10.1002/9780470559277.ch100059

Current Protocols in Chemical Biology

Current Protocols in Chemical Biology

How to Cite

Tan, E., Almaraz, R. T., Khanna, H. S., Du, J. and Yarema, K. J. 2010. Experimental Design Considerations for In Vitro Non-Natural Glycan Display via Metabolic Oligosaccharide Engineering. Current Protocols in Chemical Biology. 2:171–194.

Author Information

  1. Department of Biomedical Engineering, The Johns Hopkins University, Baltimore, Maryland

Publication History

  1. Published Online: 1 SEP 2010
  2. Published Print: SEP 2010

Abstract

Metabolic oligosaccharide engineering (MOE) refers to a technique where non-natural monosaccharide analogs are introduced into living biological systems. Once inside a cell, these compounds intercept a targeted biosynthetic glycosylation pathway and in turn are metabolically incorporated into cell-surface-displayed oligosaccharides where they can modulate a host of biological activities or be exploited as “tags” for bio-orthogonal and chemoselective ligation reactions. Undertaking a MOE experiment can be a daunting task based on the growing repertoire of analogs now available and the ever increasing number of metabolic pathways that can be targeted; therefore, a major emphasis of this article is to describe a general approach for analog design and selection and then provide protocols to ensure safe and efficacious analog usage by cells. Once cell-surface glycans have been successfully remodeled by MOE methodology, the stage is set for probing changes to the myriad cellular responses modulated by these versatile molecules. Curr. Protoc. Chem. Biol. 2:171-194 © 2010 by John Wiley & Sons, Inc.

Keywords:

  • sialic acid glycoengineering;
  • glycosylation;
  • ManNAc analogs;
  • selectin-based adhesion