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Optimal operation of high-pressure homogenization for intracellular product recovery

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Abstract

An optimal control methodology for the homogenization of bacterial cells to recover intracellular products is presented. A Fluent computational fluid dynamics (CFD) model is used to quantify the hydrodynamic forces present in the homogenizer, and empirical models are used to relate these forces to experimentally obtained cell disruption and product recovery data. The optimal homogenizer operation, in terms of either constant cell breakage or maximum intracellular product recovery, is determined using these empirical models. We illustrate this methodology with an Escherichia coli bacterial system used to produce DNA plasmids. Homogenization is performed using an industrial APV–Gaulin high-pressure homogenizer. The modeling and optimization results for this E. coli–DNA plasmid system show good agreement with the experimental data.

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Abbreviations

d :

Impact distance (mm)

f c :

Total fraction of broken cells

f f :

Fraction of cell breakage due to channel forces

f τ :

Fraction of cell breakage due to shear stress

f r :

Fraction of cell breakage due to impact ring impingement

g f :

Plasmid in the homogenizer feed pellet (g)

g h :

Plasmid in the homogenate pellet (g)

h :

Gap space (μm)

L :

Channel length (m)

P :

Operating pressure (psig)

P r :

Impact ring pressure (psig)

Q :

Volumetric flow rate (ml/s)

r p :

Fraction of free plasmid recovered from homogenization

β :

Proportionality constant

ΔE:

Post-channel turbulence energy dissipation rate (m/s)

ΔPc:

Pressure drop across the channel (psi)

ΔPi:

Channel inlet pressure gradient (Pa/m)

μ :

Viscosity (cp)

ρ :

Fluid density (g/ml)

τ m :

Maximum channel shear stress (psi)

τ w :

Fully developed channel wall shear stress (psi)

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Acknowledgements

We would like to thank APV–Gaulin for providing technical and financial support for this work. We would also like to acknowledge the contributions of Justin Miller, Mark Rogowski, and Mike Clemson in obtaining the Fluent simulation and experimental homogenizer results in this paper.

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Authors and Affiliations

  1. Department of Chemical Engineering, Villanova University, Villanova, PA, 19085-1681, USA

    William J. Kelly & Kenneth R. Muske

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  1. William J. Kelly
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  2. Kenneth R. Muske
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Correspondence to William J. Kelly.

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Kelly, W.J., Muske, K.R. Optimal operation of high-pressure homogenization for intracellular product recovery. Bioprocess Biosyst Eng 27, 25–37 (2004). https://doi.org/10.1007/s00449-004-0378-9

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  • DOI: https://doi.org/10.1007/s00449-004-0378-9

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