Glycan Variability on a Recombinant IgG Antibody Transiently Produced in HEK-293E Cells

N Biotechnol. 2012 May 15;29(4):471-6. doi: 10.1016/j.nbt.2012.02.003. Epub 2012 Mar 6.

Abstract

In this study, a recombinant monoclonal IgG antibody was produced by transient gene expression (TGE) in suspension-adapted HEK-293E cells. The objective of the study was to determine the variation in recombinant IgG yield and glycosylation in ten independent transfections. In a ten-day batch process, the variation in transient IgG yield in the ten batches was less than 30% with the specific productivity averaging 20.2 ± 2.6 pg/cell/day. We characterized the N-glycosylation profile of each batch of affinity-purified IgG by intact protein and bottom-up mass spectrometry. Four major glycans were identified at Asn(297) in the ten batches with the maximum relative deviation for a single glycoform being 2.5%. In addition, within any single transfection there was little variation in glycoforms over the ten-day culture. Our experimental data indicate that with TGE, the production of recombinant IgG with little batch-to-batch variation in volumetric yield and protein glycosylation is feasible, even in a non-instrumented cultivation system as described here.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Chromatography, High Pressure Liquid
  • Glycopeptides / analysis
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / chemistry*
  • Immunoglobulin G / genetics
  • Polysaccharides / chemistry*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Glycopeptides
  • Immunoglobulin G
  • Polysaccharides
  • Recombinant Proteins