University of Natural Resources and Life Sciences, Vienna (BOKU) - Research portal
Gewählte Master / Diploma Thesis:
Andreas Daxbacher
(2018):
Evaluation of performance of particulate Protein A affinity resins.
Master / Diploma Thesis - Institut für Biotechnologie,
BOKU-Universität für Bodenkultur,
pp 80.
UB BOKU
obvsg
FullText
- Abstract:
- The influence of velocity on process performance was investigated for two particulate Protein A affinity resins. MabSelect PrismA from GE Healthcare is a resin based on high-flow agarose and Toyopearl AF-rProtein A HC-650F has a polymethacrylate backbone. Both resins have recombinant Protein A ligands which are resistant to alkaline sanitization. The observed influence of velocity on dynamic binding capacity was expected and is also well documented in literature. In this thesis varying velocities were also tested for washing and elution steps with respect to host cell protein content and aggregation of a monoclonal antibody. However, this influence was rather small for the Toyopearl resins and negligible for MabSelect PrismA. Antibody aggregation could not be determined under any process conditions, even not for very harsh elution conditions. For Toyopearl, higher host cell protein content in eluates was determined when the column was operated at low protein saturation or higher velocities, which suggests interactions of host cell proteins primarily with the backbone. A washing step with a buffer solution containing isopropanol and urea yielded extremely low impurity levels for Toyopearl while it was only moderately effective for MabSelect PrismA. However, the impact of velocity on this removal was very small. Overall, the results of this study suggest that operating Protein A affinity media at high velocity does not affect host cell protein removal whereat the composition of the washing buffer is more important. These results are important in view of the new Protein A affinity membrane, which is currently under development at Sartorius.
- Beurteilende(r): Hahn Rainer