Ultra scale-down approach for the prediction of full-scale recovery of ovine polycolonal immunoglobulins used in the manufacture of snake venom-specific Fab fragment
Article first published online: 22 NOV 2002
Copyright © 2002 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 81, Issue 2, pages 149–157, 20 January 2003
How to Cite
Neal, G., Christie, J., Keshavarz-Moore, E. and Shamlou, P. A. (2003), Ultra scale-down approach for the prediction of full-scale recovery of ovine polycolonal immunoglobulins used in the manufacture of snake venom-specific Fab fragment. Biotechnol. Bioeng., 81: 149–157. doi: 10.1002/bit.10454
- Issue published online: 22 NOV 2002
- Article first published online: 22 NOV 2002
- Manuscript Accepted: 11 JUN 2002
- Manuscript Received: 28 FEB 2002
- the EPSRC and Protherics UK Ltd. (to G. Neal)
- ultra scale down;
- process modelling
In this article, we describe a new approach that allows the prediction of the performance of a large-scale integrated process for the primary recovery of a therapeutic antibody from an analysis of the individual unit operations and their interactions in an ultra scale-down mimic of the process. The recovery process consisted of four distinct unit operations. Using the new approach we defined the important engineering parameters in each operation that impacted the overall recovery process and in each case verified its effect by a combination of modelling and experimentation. Immunoglobulins were precipitated from large volumes of dilute blood plasma and the precipitated flocs were recovered by centrifugal separation from the liquor containing contaminating proteins, including albumin. The fluid mechanical forces acting on the precipitate and the time of exposure to these forces were used to define a time-integrated fluid stress. This was used as a scaling factor to predict the properties of the precipitated flocs at large scale. In the case of centrifugation, the performance of a full-scale disc stack centrifuge was predicted. This was achieved from a computational fluid dynamics (CFD) analysis of the flow field in the centrifuge coupled with experimental data obtained from the precipitated immunoglobulin flocs using the scale-down precipitation tank, a rotating shear device, and a standard swing-out rotor centrifuge operating under defined conditions. In this way, the performance of the individual unit operations, and their linkage, was successfully analysed from a combination of modelling and experiments. These experiments required only millilitre quantities of the process material. The overall performance of the large-scale process was predicted by tracking the changes in physical and biological properties of the key components in the system, including the size distribution of the antibody precipitates and antibody activity through the individual unit operations in the ultra scale-down process flowsheet. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 149–157, 2003.