Tailoring N-Glycan Biosynthesis for Production of Therapeutic Proteins in Saccharomyces cerevisiae

Methods Mol Biol. 2019:1923:227-241. doi: 10.1007/978-1-4939-9024-5_10.

Abstract

The ability to control and adjust the N-glycosylation pathway of Saccharomyces cerevisiae is a key step toward production of therapeutic glycoproteins such as antibodies or erythropoietin. The focus of this chapter is to describe the road from yeast-type N-glycosylation to human-type complex N-glycosylation. The chapter describes the cell engineering and provides the detailed analytical procedures required to perform glycan analysis using MALDI-TOF mass spectrometry.

Keywords: Glycan analysis; Glycoengineering; Glycosylation efficiency; Glycosyltransferases; MALDI-TOF MS; S. cerevisiae.

MeSH terms

  • Glycoproteins / genetics
  • Glycoproteins / metabolism
  • Glycosylation
  • Glycosyltransferases / genetics
  • Glycosyltransferases / metabolism
  • Polysaccharides / biosynthesis*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Glycoproteins
  • Polysaccharides
  • Saccharomyces cerevisiae Proteins
  • Glycosyltransferases