Publikationsserver der RWTH Aachen University
Yeast and plant cells as biofactories for recombinant proteins : evaluation of novel expression and downstream processing strategies
Abstract
dc:descriptionIn this thesis, the applicability of several novel developments in protein purification and analysis technologies – in particular immobilized metal ion affinity chromatography (IMAC), expanded bed adsorption (EBA) and biomolecular interaction analysis (BIA) – for recombinant protein production in yeast and tobacco was evaluated. These technologies were successfully combined into strategies for the efficient isolation of three His6-tagged recombinant proteins (FSH-specific scFv4813, CEA N-A3 hybrid protein and CEA-specific scFvT84.66) from crude yeast fermentation broth or plant cell extract, with special emphasis on scalability and cost-effectiveness. Expanded bed IMAC was successfully introduced as a novel method for capture of recombinant proteins from plant cell extract. On-line monitoring of recombinant protein purification from P.pastoris fermentation broth by real-time BIA significantly reduced overall process times. The purified proteins were analysed with respect to purity, integrity and reactivity. The aims of this thesis were achieved by the expression and downstream processing of recombinant therapeutic proteins from P.pastoris fermentation broth and N.tabacum BY-2 suspension cells. These results demonstrate the use of yeast and plant cells as biofactories for the production of therapeutic and diagnostic proteins as an important step towards the commercial exploitation of Molecular Farming.
Degree
thesis:*- Grantor dc:publisher
- Publikationsserver der RWTH Aachen University
- Year dc:date
- 2000
Author and committee
dc:creator, dc:contributor.*- Author dc:creator
-
- Drossard, Jürgen
- Contributors dc:contributor
-
- Kreuzaler, Fritz
Subjects
dc:subject × 10Rights
dc:rights- Statement dc:rights
-
- info:eu-repo/semantics/openAccess
- Language dc:language
- eng