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A comprehensive comparison of mixing, mass transfer, Chinese hamster ovary cell growth, and antibody production using Rushton turbine and marine impellers

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Abstract

Large scale production of monoclonal antibodies has been accomplished using bioreactors with different length to diameter ratios, and diverse impeller and sparger designs. The differences in these physical attributes often result in dissimilar mass transfer, mechanical stresses due to turbulence and mixing inside the bioreactor that may lead to disparities in cell growth and antibody production. A rational analysis of impeller design parameters on cell growth, protein expression levels and subsequent antibody production is needed to understand such differences. The purpose of this study was to examine the impact of Rushton turbine and marine impeller designs on Chinese hamster ovary (CHO) cell growth and metabolism, and antibody production and quality. Experiments to evaluate mass transfer and mixing characteristics were conducted to determine if the nutrient requirements of the culture would be met. The analysis of mixing times indicated significant differences between marine and Rushton turbine impellers at the same power input per unit volume of liquid (P/V). However, no significant differences were observed between the two impellers at constant P/V with respect to oxygen and carbon dioxide mass transfer properties. Experiments were conducted with CHO cells to determine the impact of different flow patterns arising from the use of different impellers on cell growth, metabolism and antibody production. The analysis of cell culture data did not indicate any significant differences in any of the measured or calculated variables between marine and Rushton turbine impellers. More importantly, this study was able to demonstrate that the quality of the antibody was not altered with a change in the impeller geometry.

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Abbreviations

CHO:

Chinese hamster ovary

P :

Power (W)

V :

Volume (L)

DHFR:

Dihydrofolate reductase

Re :

Reynolds number (dimensionless)

IgG:

Immunoglobulin

HPLC:

High-performance liquid chromatography

H :

Liquid height in reactor (m)

T :

Reactor diameter (m)

DO:

Dissolved oxygen (%)

ρ :

Density (kg/m3)

P 0 :

Power number (marine 0.4, Rushton 5)

N :

Agitation rate (rev/s)

D :

Impeller diameter (m)

n :

Number of impellers

μ :

Viscosity (mPa s)

θ m :

Mixing time (s)

ε T :

Mean specific energy dissipation rate (W/kg)

k L a O :

Mass transfer coefficient for oxygen in culture medium (h−1)

k L a sO :

Surface mass transfer coefficient for oxygen (h−1)

vvh:

Volume of gas per volume of liquid per hour

slpm:

Standard liters per minute

ppm:

Parts per million

C*:

Saturation concentration of oxygen (% dissolved oxygen)

C L :

Concentration of oxygen gas in bulk liquid (% dissolved oxygen)

OUR:

Oxygen utilization rate (% dissolved oxygen/h)

C oxygen :

Concentration of oxygen (% dissolved oxygen)

C T :

Total inorganic carbon

k L a C :

Mass transfer coefficient for carbon dioxide (h−1)

k corr :

Correction factor (dimensionless)

ANOVA:

Analysis of variance

X :

Total cell concentration (cells mL−1)

X V :

Viable cell concentration (cells mL−1)

μ :

Specific growth rate of cells (day−1)

μ max :

Maximum specific growth rate of cells (day−1)

q P :

Specific product formation rate (pg/cell/day)

α :

Growth associated product rate constant

β :

Non-growth associated product rate constant

q Ab :

Specific rate of antibody production (pg/cell/day)

q L :

Specific rate of lactate production (pg/cell/day)

q N :

Specific rate of ammonia production (pg/cell/day)

CFD:

Computational fluid dynamics

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Author notes

  1. Sandeepa Sandadi

    Present address: BioPharmaceutical Development, Human Genome Sciences, Inc., 14200 Shady Grove Road, Rockville, MD, 20850, USA

Authors and Affiliations

  1. BioProcess Development, Merck Research Laboratories, 1011 Morris Avenue, Union, NJ, 07083, USA

    Sandeepa Sandadi, John S. Bowers & Dennis Rendeiro

  2. Department of Chemical and Biochemical Engineering, Rutgers, The State University of New Jersey, 98 Brett Road, Piscataway, NJ, 08854, USA

    Henrik Pedersen

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  1. Sandeepa Sandadi
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  2. Henrik Pedersen
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  4. Dennis Rendeiro
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Correspondence to Sandeepa Sandadi.

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Sandadi, S., Pedersen, H., Bowers, J.S. et al. A comprehensive comparison of mixing, mass transfer, Chinese hamster ovary cell growth, and antibody production using Rushton turbine and marine impellers. Bioprocess Biosyst Eng 34, 819–832 (2011). https://doi.org/10.1007/s00449-011-0532-0

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  • DOI: https://doi.org/10.1007/s00449-011-0532-0

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