Glutamine-free mammalian expression of recombinant glycoproteins with uniform isotope labeling: an application for NMR analysis of pharmaceutically relevant Fc glycoforms of human immunoglobulin G1

J Biomol NMR. 2022 Apr;76(1-2):17-22. doi: 10.1007/s10858-021-00387-5. Epub 2022 Jan 3.

Abstract

Mammalian cells are widely used for producing recombinant glycoproteins of pharmaceutical interest. However, a major drawback of using mammalian cells is the high production costs associated with uniformly isotope-labeled glycoproteins due to the large quantity of labeled L-glutamine required for their growth. To address this problem, we developed a cost-saving method for uniform isotope labeling by cultivating the mammalian cells under glutamine-free conditions, which was achieved by co-expression of glutamine synthase. We demonstrate the utility of this approach using fucosylated and non-fucosylated Fc glycoforms of human immunoglobulin G1.

Keywords: Fucose; Glutamine synthase; Glycoprotein; Mammalian expression system; Metabolic isotope labeling; Therapeutic antibody.

MeSH terms

  • Animals
  • Glutamine*
  • Glycoproteins* / chemistry
  • Humans
  • Immunoglobulins
  • Isotope Labeling / methods
  • Mammals
  • Nuclear Magnetic Resonance, Biomolecular
  • Recombinant Proteins / chemistry

Substances

  • Glycoproteins
  • Immunoglobulins
  • Recombinant Proteins
  • Glutamine