Adapted ultra scale-down approach for predicting the centrifugal separation behavior of high cell density cultures

Andrew D. Tustian, Heidi Salte, Nicholas A. Willoughby, Inass Hassan, Michael H. Rose, Frank Baganz, Michael Hoare, Nigel J. Titchener-Hooker

Research output: Contribution to journalArticle

Abstract

The work presented here describes an ultra scale-down (USD) methodology for predicting centrifugal clarification performance in the case of high cell density fermentation broths. Existing USD approaches generated for dilute systems led to a 5- to 10-fold overprediction of clarification performance when applied to such high cell density feeds. This is due to increased interparticle forces, leading to effects such as aggregation, flocculation, or even blanket sedimentation, occurring in the low shear environment of a laboratory centrifuge, which will not be apparent in the settling region of a continuous-flow industrial centrifuge. A USD methodology was created based upon the dilution of high solids feed material to ~2% wet wt/vol prior to the application of the clarification test. At this level of dilution cell-cell interactions are minimal. The dilution alters the level of hindered settling in the feed suspensions, and so mathematical corrections are applied to the resultant clarification curves to mimic the original feed accurately. The methodology was successfully verified: corrected USD curves accurately predicted pilot-scale clarification performance of high cell density broths of Saccharomyces cerevisiae and Escherichia coli cells. The USD method allows for the rapid prediction of large-scale clarification of high solids density material using millilitre quantities of feed. The advantages of this method to the biochemical engineer, such as the enabling of rapid process design and scale-up, are discussed. © 2007 American Chemical Society and American Institute of Chemical Engineers.

Original languageEnglish
Pages (from-to)1404-1410
Number of pages7
JournalBiotechnology Progress
Volume23
Issue number6
DOIs
Publication statusPublished - Nov 2007

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Cell culture
Cells
Dilution
Centrifuges
Engineers
Flocculation
Sedimentation
Yeast
Fermentation
Escherichia coli
Process design
Suspensions
Agglomeration

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Tustian, Andrew D. ; Salte, Heidi ; Willoughby, Nicholas A. ; Hassan, Inass ; Rose, Michael H. ; Baganz, Frank ; Hoare, Michael ; Titchener-Hooker, Nigel J. / Adapted ultra scale-down approach for predicting the centrifugal separation behavior of high cell density cultures. In: Biotechnology Progress. 2007 ; Vol. 23, No. 6. pp. 1404-1410.
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abstract = "The work presented here describes an ultra scale-down (USD) methodology for predicting centrifugal clarification performance in the case of high cell density fermentation broths. Existing USD approaches generated for dilute systems led to a 5- to 10-fold overprediction of clarification performance when applied to such high cell density feeds. This is due to increased interparticle forces, leading to effects such as aggregation, flocculation, or even blanket sedimentation, occurring in the low shear environment of a laboratory centrifuge, which will not be apparent in the settling region of a continuous-flow industrial centrifuge. A USD methodology was created based upon the dilution of high solids feed material to ~2{\%} wet wt/vol prior to the application of the clarification test. At this level of dilution cell-cell interactions are minimal. The dilution alters the level of hindered settling in the feed suspensions, and so mathematical corrections are applied to the resultant clarification curves to mimic the original feed accurately. The methodology was successfully verified: corrected USD curves accurately predicted pilot-scale clarification performance of high cell density broths of Saccharomyces cerevisiae and Escherichia coli cells. The USD method allows for the rapid prediction of large-scale clarification of high solids density material using millilitre quantities of feed. The advantages of this method to the biochemical engineer, such as the enabling of rapid process design and scale-up, are discussed. {\circledC} 2007 American Chemical Society and American Institute of Chemical Engineers.",
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Tustian, AD, Salte, H, Willoughby, NA, Hassan, I, Rose, MH, Baganz, F, Hoare, M & Titchener-Hooker, NJ 2007, 'Adapted ultra scale-down approach for predicting the centrifugal separation behavior of high cell density cultures', Biotechnology Progress, vol. 23, no. 6, pp. 1404-1410. https://doi.org/10.1021/bp070175d

Adapted ultra scale-down approach for predicting the centrifugal separation behavior of high cell density cultures. / Tustian, Andrew D.; Salte, Heidi; Willoughby, Nicholas A.; Hassan, Inass; Rose, Michael H.; Baganz, Frank; Hoare, Michael; Titchener-Hooker, Nigel J.

In: Biotechnology Progress, Vol. 23, No. 6, 11.2007, p. 1404-1410.

Research output: Contribution to journalArticle

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