Transient expression of an IL-23R extracellular domain Fc fusion protein in CHO vs. HEK cells results in improved plasma exposure

Protein Expr Purif. 2010 May;71(1):96-102. doi: 10.1016/j.pep.2009.12.015. Epub 2010 Jan 4.

Abstract

Transient transfection of mammalian cells in suspension culture has recently emerged as a very useful method for production of research-scale quantities of recombinant proteins. The most commonly used cell lines for this purpose are suspension-adapted HEK and CHO cells. We report here that the plasma exposure in mice of an IL-23R extracellular domain Fc fusion protein (IL23R-Fc) differed dramatically depending on whether the protein was prepared by transient transfection of HEK or CHO cells. Specifically, IL23R-Fc expressed using CHO cells had about 30-fold higher in vivo plasma exposure compared to the HEK-expressed protein. In contrast to their differing plasma exposures, the HEK- and CHO-expressed proteins had equivalent in vitro biological activity. Characterization of the CHO- and HEK-expressed IL23R-Fc proteins indicated that the differences in in vivo plasma exposure between them are due to differential glycosylation.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Electrophoresis, Polyacrylamide Gel
  • Extracellular Space / metabolism*
  • Glycosylation
  • Humans
  • Isoelectric Focusing
  • Lectins / metabolism
  • Mice
  • Molecular Sequence Data
  • Protein Structure, Tertiary
  • Receptors, Fc / metabolism*
  • Receptors, Interleukin / blood*
  • Receptors, Interleukin / chemistry*
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism*
  • Recombinant Fusion Proteins / pharmacokinetics
  • Transfection / methods*

Substances

  • IL23R protein, human
  • Lectins
  • Receptors, Fc
  • Receptors, Interleukin
  • Recombinant Fusion Proteins