Research

Molecular Biotechnology

, Volume 34, Issue 2, pp 191-199

First online:

Serum-free suspensin large-scale transient transfection of CHO cells in WAVE bioreactors

  • Raj HaldankarAffiliated withDepartment of Protein Sciences, Amgen, Inc. Email author 
  • , Danqing LiAffiliated withDepartment of Protein Sciences, Amgen, Inc.
  • , Zane SaremiAffiliated withDepartment of Protein Sciences, Amgen, Inc.
  • , Claudia BaikalovAffiliated withDepartment of Protein Sciences, Amgen, Inc.
  • , Rohini DeshpandeAffiliated withDepartment of Protein Sciences, Amgen, Inc.

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Abstract

Here, we report the development of a large-scale transient expression platform utilizing Chinese hamster ovary (CHO) cells. The majority of recombinant proteins and antibodies that are produced for preclinical models and clinical trials are expressed in stably transfected CHO cells. A protocol for transient transfection of CHO cells that is rapid, reproducible, and cost-effective would therefore streamline the process from research to development and help avoid any potential host species induced variation in the molecule of interest. CHO cells were adapted to grow in serum-free suspension conditions in spinner flask cultures in a proprietary in-house developed growth medium. In developing this transient transfection protocol, the parameters optimized included the transfection reagent of choice, the cell density at the time of transfection, the plasmid DNA concentration, and the transfection reagent concentration. Using this optimized protocol, we have expressed recombinant proteins, including antibodies, at an expression level of up to 9.4 mg/L. We also report transient transfections from 500 mL working volume (w.v.) up to 20 L w.v. in a WAVETM bioreactor. Using this optimized protocol, it is possible to rapidly (within 10 d) produce up to 100 mg of recombinant protein for further study.

Index Entries

Transient transfection CHO cells WAVE bioreacter polyethyleneimine recombinant protein expression