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BioMetals

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Nickel and cobalt affect galactosylation of recombinant IgG expressed in CHO cells

  • Anuja Prabhu
  • Mugdha Gadgil
Article
  • 143 Downloads

Abstract

Glycosylation is an important product quality attribute of antibody biopharmaceuticals. It involves enzymatic addition of oligosaccharides on proteins by sequential action of glycosyltransferases and glycosidases in the endoplasmic reticulum and golgi. Some of these enzymes like galactosyltransferase and N-acetylglucosaminyltransferase-I require trace metal cofactors. Variations in trace metal availability during production can thus affect glycosylation of recombinant glycoproteins such as monoclonal antibodies. Variability in trace metal concentrations can be introduced at multiple stages during production such as due to impurities in raw materials for culture medium and leachables from bioreactors. Knowledge of the effect of various trace metals on glycosylation can help in root-cause analysis of unintended variability in glycosylation. In this study, we investigated the effect of nickel and cobalt on glycosylation of recombinant IgG expressed in Chinese hamster ovary cells. Nickel concentrations below 500 µM did not affect glycosylation, but above 500 µM it significantly decreases galactosylation of IgG. Cobalt at 50 µM concentration causes slight increase in G1F glycans (mono galactosylated) as previously reported. However, higher concentrations result in a small increase in G0F (non galactosylated) glycans. This effect of nickel and cobalt on galactosylation of recombinant IgG can be reversed by supplementation of uridine and galactose which are precursors to UDP-Galactose, a substrate for the enzymatic galactosylation reaction.

Keywords

Glycosylation Trace metals Process variability Nickel Cobalt Galactosylation 

Notes

Acknowledgements

MG acknowledges funding from the Department of Biotechnology, Government of India. The authors are thankful to the MALDI-MS facility at CSIR-NCL and to Dr. Gadre for help with HPLC-based galactosyltransferase assay.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10534_2018_152_MOESM1_ESM.docx (17 kb)
Supplementary material 1 (DOCX 17 kb)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Chemical Engineering and Process Development DivisionCSIR-National Chemical LaboratoryPuneIndia

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