title: Experimental Design Considerations for In Vitro Non-Natural Glycan Display via Metabolic Oligosaccharide Engineering.
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.
authors.affiliation: Department of Biomedical Engineering, The Johns Hopkins University, Baltimore, Maryland.
authors.forename: Elaine
authors.initials: E
authors.lastname: Tan
authors.forename: Ruben T
authors.initials: RT
authors.lastname: Almaraz
authors.forename: Hargun S
authors.initials: HS
authors.lastname: Khanna
authors.forename: Jian
authors.initials: J
authors.lastname: Du
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Yarema
keywords: ManNAc analogs; glycosylation; selectin‐based adhesion; sialic acid glycoengineering