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Suppression of programmed cell death in industrial scale biological production systems

   
Project

QLK3-2000-00076

Cell factory area

3.1.2

EU Contribution

884 161 Euro

Duration

36 months

Type

Demonstration project

Starting date

01-01-2001

Keywords
anti-apoptosis genes
bcl-2
programmed cell death
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ABSTRACT
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Apoptosis is the genetically determined form of controlled cellular elimination that accounts for most of the cell death that takes place in vivo. It is also the primary mechanism of cell death during the in vitro cultivation of mammalian cell lines used in the production of biopharmaceuticals. In recent years, lab scale studies have indicated that the suppression of the apoptotic pathway during the synthesis of these high value products can substantially increase the productivity of the culture process. In the demonstration proposed here, we will develop new cell lines that incorporate anti-apoptosis genes and will demonstrate the impact of this strategy on the efficiency of state-of-the-art industrial-scale biopharmaceutical production processes, and then make them available to the biotechnology industry.

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OBJECTIVES
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  • To assess the impact of the bcl-2 gene on process development, fermentation and product recovery under state-of-the-art biopharmaceutical manufacturing conditions.
  • To demonstrate the impact of bcl-2 expression on cellular productivity in pilot-scale perfusion cultures and during the development of feeding strategies for fed-batch cultures.
  • To demonstrate that bcl-2 reduces the rate of cell death in a range of genetic backgrounds by examining the effect on a panel of NSO and CHO clones.
  • To construct fully characterised and validated "parental" cell lines over-expressing bcl-2 and to allow exploitation of these cell lines by the industrial partner to be used as hosts for the production of biopharmaceuticals.

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DESCRIPTION OF THE WORK
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At lab scale, the suppression of apoptosis has been shown to result in significant improvements in culture reproducibility, enhanced adaptation to protein-free conditions, and increase product titres. The proposal described here will represent the next key step in the development of this approach for the optimisation of industrial cell cultures: the demonstration of the impact of bcl-2 on the economics of the manufacturing process. The impact of enhanced cellular robustness on cloning and selection of over-producers will be assessed, as will the speed of adaptation to protein-free media. The effect of increased robustness will be analysed during the inoculum scale-up and fermentation processes, with all culture parameters being closely monitored. The effect of improved culture viability on product recovery will also be determined. This phase of the study will provide a complete picture with regard to the impact of bcl-2 on process economics. We will compare the impact of bcl-2 on NSO and CHO growth rate, apoptosis and productivity in a variety of culture systems, including batch, fed batch, perfusion and continuous, from bench-top through to pilot and ultimately under state of the art industrial scale conditions.

The use of bcl-2 cells will also be assessed when developing novel feeding strategies during fed-batch cultures. In addition, it will be important to find out whether bcl-2 can protect different NSO and CHO cell lines from apoptosis, to show the reproducibility of this approach. CHO and NSO cell lines developed during this stage of the project will then be validated and commercially exploited by the partners.

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DELIVERABLES
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  • The project will demonstrate that the over-expression of anti-apoptosis genes provides many advantages during bioprocess development, fermentation and product recovery.
  • It will demonstrate that the use of highly robust cell lines raises the overall productivity of the system, resulting in increased profitability and speed with which the biopharmaceuticals can be made available to clinicians.
  • It will also generate a bank of highly robust cell lines, which will be commercialised by the partners.

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CONSORTIUM
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COORDINATOR
  Dr. Mohamed Al-Rubeai
University of Birmingham
School of Chemical Engineering
B15 2TT Birmingham, United Kingdom
Tel: +44 1214143888
Fax: +44 1214143888
m.al-rubeai@bham.ac.uk

PARTNERS
  Dr. Ralf Portner
Technische Universitat Hamburg Harburg
21071 Hamburg, Germany
Tel: +49 40428782886
Fax: +49 40428782909
poertner@tu-hamburg.de

Prof. John Birch
Lonza Biologics PLC
Research and Development
SL1 4DY Slough, United Kingdom
Tel: +44 1753777000
Fax: +44 1753777001
jbirch@lonza.co.uk
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