Fraunhofer IME Master Class - Master Thesis offer at Fraunhofer IME in Aachen, in the Department Bioprocess engineering
Title: Expression and Purification of Tobacco Etch Virus protease in Nicotiana benthamiana its chromatographic characterization

Abstract
Tobacco Etch Virus (TEV) Protease is a highly specific cysteine protease often used for the removal of affinity tags and mutations were introduced to decrease autolysis. Commercially available TEV Protease is mostly expressed in Escherichia coli. However, the expression of TEV protease in Nicotiana species is promising due to the Tobacco Etch Virus origin. Another interesting aspect is the application of TEV protease as a model protein to characterize chromatography. Development of classical chromatography methods like ion exchange chromatography or hydrophobic interaction chromatography takes allot of effort and time. In order to reduce the number of experiments, we want to set-up a model for elution prediction. Part of this model is the steric mass action (SMA) model, which describes multicomponent protein-salt equilibria in ion-exchange systems, facilitating predictions about adsorption and desorption reactions.

Task
The student accepted for the project/thesis described above will first have to express TEV protease in Nicotiana benthamina and purify sufficient amounts for SMA parameter determination (at least 5 mg). To do so, the student will have to clone the vector in N. benthamiana, carry out infiltration with Agrobacterium tumefaciens and finally extract, clarify and purify TEV from infiltrated plants, analyze them by SDS-PAGE, capillary gel electrophoresis, Bradford assay and additional protein quantification methods. The SMA parameter should be determined for at least 3 different ion exchange columns.

Laboratory skills
(S)He has good basic knowledge in biochemistry and bioengineering and is willing to gain more insights into the issues faced by today’s plant and pharmaceutical biotechnology. At least basic experience in downstream processing is mandatory for a successful application.

Language skills
Fluent english and/or german

Computer skills
Excel and Powerpoint

Timeline
Earliest starting date 01.06.2021; expected Duration (months): > 6

Supervisor contact information
Name: Catherine Bernau
E-Mail: Catherine.bernau@ime.fraunhofer.de
Phone: + 49 241 6085-35001


In case of identical qualifications preference will be given to severely disabled candidates.
We would like to point out that the chosen job title also includes the third gender.
The Fraunhofer-Gesellschaft emphasises gender-independent professional equality.
Job Reference: IME-MB-2021-9 Closing Date: 30.06.2021
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