Production of recombinant h-AT III with mammalian cell cultures using capillary electrophoresis for product monitoring.

Cytotechnology
R FreitagG Kretzmer

Abstract

The importance of mammalian cell cultures for biotechnological production processes is steadily increasing, despite the high demands of these organisms on their culture conditions. Efforts towards a more efficient bioprocess generally concentrate on maximizing the culture's life time, the cell number, and the product concentration. Here recombinant BHK 21 c13 cells are used to produce rh-AT III, an anticoagulant of high therapeutic value. The influence of the process mode (batch, repeated batch, continuous perfusion) and the process temperature (30°C vs. 37°C) on the above mentioned parameters is investigated. It is possible to increase the length of the culture from 140 h (batch) to more than 500 h (continuous perfusion culture), while concomitantly increasing the cell density from 0.72 10(6)/ml (batch) to 2.27 10(6)/ml (repeated batch) and 2.87 10(6)/ml (continuous perfusion culture). The accumulation of toxic metabolites, such as lactate, can be curtailed by reducing the bioreactor temperature from 37°C to 30°C during the later part of the exponential growth phase. Fast and reliable product monitoring became essential during process optimization. Capillary zone electrophoresis (CZE) in uncoated fused silica capillaries was s...Continue Reading

References

Jan 1, 1992·Enzyme and Microbial Technology·D Wentz, K Schügerl
Sep 11, 1992·Journal of Chromatography·A VintherH Søeberg
Aug 1, 1990·Biochimie·L MoureyD Moras
Aug 11, 1989·Journal of Chromatography·B L KargerA Guttman
Jan 1, 1983·Annual Review of Biochemistry·J Travis, G S Salvesen
Jan 1, 1994·Cytotechnology·R WeidemannG Kretzmer
Nov 12, 1993·Journal of Chromatography. a·R LauschR Freitag

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Citations

Dec 22, 1999·Electrophoresis·A L Lagu

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