Fraunhofer Institute for Molecular Biology and Applied Ecology

3 ARTICLES PUBLISHED IN JoVE

Bioengineering

Characterization of Complex Systems Using the Design of Experiments Approach: Transient Protein Expression in Tobacco as a Case Study

Johannes Felix Buyel¹, Rainer Fischer^1,2

¹Institute for Molecular Biotechnology, RWTH Aachen University, ²Institute for Molecular Biology and Applied Ecology, Fraunhofer Gesellschaft

We describe a design of experiments approach that can be used to determine and model the influence of transgene regulatory elements, plant growth and development parameters, and incubation conditions on the transient expression of monoclonal antibodies and reporter proteins in plants.

Environment

Expression of Recombinant Cellulase Cel5A from Trichoderma reesei in Tobacco Plants

Megan Garvey¹, Johannes Klinger^*1, Holger Klose^*1, Rainer Fischer², Ulrich Commandeur¹

¹Institute for Molecular Biotechnology, Bio7, RWTH Aachen University, ²Fraunhofer Institute for Molecular Biology and Applied Ecology

Tobacco plants were used to produce a fungal cellulase, TrCel5A, via a transient expression system. The expression could be monitored using a fluorescent fusion protein, and the protein activity was characterized post-expression.

Biology

Comparison of Tobacco Host Cell Protein Removal Methods by Blanching Intact Plants or by Heat Treatment of Extracts

Johannes F. Buyel^1,2, Jürgen Hubbuch³, Rainer Fischer^1,2

¹Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e. V., ²Institute for Molecular Biotechnology, RWTH Aachen University, ³Department of Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT)

Three heat precipitation methods are presented that effectively remove more than 90% of host cell proteins (HCPs) from tobacco extracts prior to any other purification step. The plant HCPs irreversibly aggregate at temperatures above 60 °C.