Large-Scale Transient Production in ExpiCHO-S™ with Enhanced N-Galactosylation-Sialylation and PEI-Based Transfection

Methods Mol Biol. 2022:2313:143-150. doi: 10.1007/978-1-0716-1450-1_7.

Abstract

Large-scale transient expression in Chinese Hamster Ovary (CHO) cells provides a rapid protein production method with a potential start-to-end alignment advantage for biotherapeutics drug discovery. In this chapter, experimental protocols are illustrated for transient expression of therapeutic glycoproteins with improved galactosylation and sialylation in ExpiCHO-S™ system. To reduce the production cost, we also describe a novel procedure for PEI-mediated transfection in ExpiCHO-S™ cells that supports therapeutic protein expression comparable to the level with ExpiFectamine™-based transfection.

Keywords: Chinese Hamster Ovary (CHO) Cells; N-linked glycosylation and sialylation; Polyethylenimine; Therapeutic glycoproteins; Transfection; Transient expression.

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Protein Processing, Post-Translational
  • Recombinant Proteins
  • Transfection*

Substances

  • Recombinant Proteins