Performance Comparison of CHO-S Cell Cultivation and mAb Production in the Allegro™ STR 200 Single-Use Bioreactor and in Conventional Single-Use Bioreactor



Pall: Odette Becheau, Emmanuelle Cameau, John Welsh, Kyle Jones, Joe Capone and Marie-Laure Collignon

ZHAW: Sören Werner , Cedric Schirmer, Nina Steffen, Jan Müller, Regine Eibl and Dieter Eibl




Today, the benefits of single-use (SU) technologies for upstream processing are much more prevalent and heavily adopted within biopharmaceutical industry. Pall has developed the Allegro™ STR stirred single-use bioreactor family which is currently available in 4 sizes: STR 50, STR 200, STR 1000, and STR 2000 (Figure 1). The direct-bottom mechanically driven impeller allows large range of power inputs from 0.002 W/kg up to 0.3 W/kg, while the macro sparger seated below the three 45-degree elephant ear blades results in high oxygen transfer rates (kLa up to 40 h-1) and short mixing time (minimum tM~10 s).


Previous studies highlighted that the critical parameters to be kept constant from one scale to another are the volumetric power input (P/V) for agitation and the superficial gas velocity (vs) for aeration. This will successfully scale up a process at constant kLa, keeping equivalent mixing time and shear stress environment in the bioreactor.






Cell Line, Medium and Supplements


  • Chinese hamster ovary (CHO)-S clone 38 cell line, producing a human IgG antibody
  • ActiPro♦ medium (GE Healthcare) supplemented with:
    • 4 mM UltraGlutamine♦ 1 (Lonza)
    • 1x Gibco♦ HT Supplement mixture (Thermo Fisher Scientific)
    • Puromycin (1.25 g/L, 10 mg/mL)
  • Feed with HyClone♦ Cell Boost 7a and Cell Boost 7b (GE Healthcare) start in day 3
  • Glucose solution (45%, 450 g/L) for feed if glucose was below 4 g/L
  • Antifoam C emulsion (Sigma Aldrich)




  • 125 and 500 mL Erlenmeyer shake flasks (Corning) for inoculum production
  • Flexsafe♦ RM wave bioreactor bag (Sartorius) for inoculum production
  • Allegro STR 200 single-use bioreactor system with heat mat (Pall)
  • Conventional single-use bioreactor


Operating parameters for the cultivation for Pall’s Allegro STR 200 single-use bioreactor and for the conventional single-use bioreactor are shown in Table 1






  • Polarographic oxygen sensor InPro♦ 6800 (Mettler-Toledo) for Allegro STR 200 single-use bioreactor
  • pH sensor (EasyFerm Hamilton) for Allegro STR 200 single-use bioreactor
  • Single-use pH and oxygen sensor incorporated in conventional single-use bioreactor
  • BioProfile♦ 100 Plus ID 2 (Nova Biomedical)
  • Cedex HiRes and Cedex Bio Analyzer (Roche)
  • Prominence HPLC System (Shimadzu)
  • Xevo♦ G2 LC-MS and Acquity H-Class Bio UPLC systems (Waters)




  • Simplified handling of a novel bioreactor system
  • Inoculum production with shake flask and wave-mixed bag is an appropriate method
  • Average peak cell density in the Allegro STR 200 single-use bioreactor at the end of the growth phase was 2.15 · 107 cells·mL-1 (Figure 2a)
  • Cell metabolism in terms of glucose uptake and lactate formation is very comparable (Figure 2b)
  • Average monoclonal antibody (mAb) titer of 2.18 g·L-1 at the culture harvest day (day 10) (Figure 2c)
  • The main glycan composition shows a similar finger print for both bioreactors flor species G0F, G1F and G2F (Figure 2d)
  • Aggregates in both systems and amount of molecular weight species (LMW) (Figure 2e)






  • The Pall Allegro STR 200 stirred single-use bioreactor demonstrated excellent performance to successfully cultivate this CHO culture
  • The large elephant ear impeller exhibits efficient mixing and effective oxygen transfer. No limitations could be detected during the cultivations
  • The profiling of mAb quality and the mAb quantity show that the single impeller's shear profile does not influence the antibody
  • Further, using one single impeller only, feeding processes can be applied with high degree of possible working volume




  1. Schirmer, Müller J., Steffen N., Werner S., Eibl R. & Eibl D., “How to produce mAbs in a cube-shaped stirred single-use bioreactor at 200 L scale”. R. Pörtner, editor. Animal Cell Biotechnology. Methods in Molecular Biology (Methods and Protocols), Fourth Edition. New York, Springer Science+Business Media, LLC (Publication forthcoming)
  2. “Characterization and engineering performance of the Allegro STR 1000 stirred single-use tank bioreactor” Pall Application Note USD 3136
  3. “Cultivation of chinese hamster ovary (CHO) cells in Allegro STR 1000 stirred single-use tank bioreactor system” Pall Application Note USD 3135



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