| Autor: |
Ewald Benes, Gordana Simic, Otto Doblhoff-Dier, Felix Trampler, Renate Kunert, Hermann Katinger, Timo Keijzer, Dethardt Müller, Martin Gröschl, Willibald Steinfellner |
| Rok vydání: |
2001 |
| Předmět: |
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| Zdroj: |
Animal Cell Technology: From Target to Market ISBN: 9789401038973 |
| DOI: |
10.1007/978-94-010-0369-8_68 |
| Popis: |
The expression of recombinant glycoproteins that bear a cytotoxic potential is a major challenge using animal cells. Product yields in a range of 0.1 g/l to 1.0g/l as commonly reached for proteins that do not adversely affect the cultured cells are out of reach. The permanent expression of a cytotoxic protein at low specific productivities requires a bioprocess that allows the accumulation of product at concentrations adequate for purification. We compared a continuous perfusion process based on an ultrasonic cell retention device with a discontinuous fed-batch using a balanced nutrient concentrate. Theproduct yield in batch mode was set to a relative concentration of 1.0. In fed-batch the product yield was increased by 40% to a relative value of 1.4 accompanied by a significant loss of culture viability indicating the detrimental effect. However, under perfused conditions not only the volumetric productivity was increased as expected, but also the relative product concentration reached a valueof 6.0 at viabilities of more than 90%. Furthermore the product was harvested almost cell free at perfusion rates of 2d−1. Combining the perfusion mode (cell accumulation) and the fed-batch mode (product accumulation) the product yield was significandy increased toa relative value of 10.0 at final viabilities of still 65%. We thus developed a process capable of accumulating a cytotoxic protein in concentrations sufficient for downstream processing requirements. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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