752347 Practical course in protein engineering and technology (in Eng.)

Art
Praktikum
Semesterstunden
5
Vortragende/r (Mitwirkende/r)
Poliak, Peter , Hofbauer, Stefan , Furtmüller, Paul Georg , Altmann, Friedrich , Ludwig, Roland , Wozniak-Knopp, Gordana , Schütz, Georg , Petrov, Drazen
Organisation
Angeboten im Semester
Sommersemester 2025
Unterrichts-/ Lehrsprachen
Englisch
Lehrinhalt

The practical course in protein engineering and technology is meant to conclude your studies within the specialization "Protein Engineering and Technology" of the Biotechnology Master (H418).

The participants of this course will work in a team to achieve their goals, which are different in the summer semester and winter semester. In the winter semester, you will be part of a (fictional) biosensor start-up that develops biosensors for medical applications, whereas in the summer semester you will join a (fictional) company research team to develop bispecific antibodies. You are encouraged to choose the topic of your interest.

Winter semester:
A (fictional) start-up wants to develop its first prototype of a blood glucose biosensor. The timeline is killing and the budget is tight. You are part of the research team that gathers information from all available sources and you plan, perform and analyze laboratory experiments. The prototype you want to develop is based on the currently most advanced biosensor platform. For your third-generation biosensor, you want to apply cellobiose dehydrogenase (CDH), a two-domain enzyme capable of direct electron transfer. Because of new FDA regulations, you will not only have to prove that your prototype can detect glucose in the blood, but also that it is not affected by maltose, which could cause patient fatalities. Based on homology modelling, docking to the active-site of CDH, predicting protein properties and effects of mutations using computational approaches you determine your candidate CDHs. Based on these results you have to verify the quality of the available CDHs by methods of protein analytics. Which CDH is the most stable (differential scanning calorimetry (DSC)), has a high cofactor loading and a native conformation (circular dichroism (CD) spectroscopy), is least glycosylated (mass spectrometry, MS), or most active (steady-state and pre-steady-state kinetics)? Finally, you will prepare biosensors with the selected CDHs and test their performance in different buffers and for different substrates. At the end you will present your research and the prototype to a consortium of investors.

Summer semester:

A small team of scientists in the research division of a big pharma company has developed a new cool bispecific antibody format and wishes to forward it for manufacturing, so they defend their invention and show its advantages among over 30 different formats available. You will use modelling approaches to predict relevant properties of the antibody (e.g. stability), its interactions to the cognate antigens and the effects of point mutations.You will determine heterodimer purity with analytical methods (SEC, MS) and its thermostability with DSC. You will examine if the binding to two cognate antigens is retained with affinity measurements via BLI, and perform a cell surface titration to verify that this still holds for a cell-bound antigen. Finally, the result of a biological cell-based assay will demonstrate the activity of the bispecific antibody. With this, you will wrap-up a data package to be presented to the senior management to support their decision on the future of your favorite project.
Inhaltliche Voraussetzungen (erwartete Kenntnisse)

Applicants are expected to have participated and finished most of the courses in the specialization of Protein Engineering and Technology, especially VU 772328 “Methods in Protein Characterization”, VU 894308 “Modelling and Simulation of Biomolecules” and VO 752345 “Enzyme Reactions: Mechanisms and Kinetics”.
Please also study the available information on BOKUlearn (Moodle) carefully before our first, introductory meeting.

You should:
- be able to download, edit and save protein sequences
- to perform basic homology modelling and use PyMol/MOE as a structure viewing program
- have understood the principles of molecular docking
- know how to use methods for protein analysis like DSC, CD, MS, SEC, Octet (biolayer interferometry)
- be trained to measure enzyme kinetics (winter semester) or binding affinity (summer semester)
- be highly motivated to collaborate with your colleagues
- participate in the management of your research team
Lehrziel

After successfully finishing the practical course in Protein Engineering and Technology you will fulfill the following criteria:

1. Knowledge: Identify amino acids in protein structures and their contribution to protein properties; Know methods to analyze protein structures and methods to analyze proteins

2. Comprehension: Explain the principles of molecular docking and the rationale for choosing different protein analysis methods

3. Application: Select the most appropriate method/approach to investigate and characterize proteins in silico and in vitro; Apply enzymes in biosensor production or antibodies in medical diagnostics

4. Analysis: Define relevant research tasks in a project; Break-down the research into manageable tasks and evaluate the performance of team members within a sub-task; Analyze obtained data and their significance for the project.

5. Synthesis: Recognize important results; Combine results to select the most suitable protein; Organize the most efficient production of the prototype

6. Evaluation: Assess the success of your project; Predict the chances of your prototype to be commercialized

Antibody Engineering Practical Course:
The successful participants of the course, future antibody engineers, will be able to:
-Implement the relevant techniques to determine the biophysical properties of bispecific antibodies and compare the results of the measurements for their asset molecule with the controls
-Interpret the outcomes of the biophysical evaluation of their asset molecule and propose possible improvements
-Design an in-vitro differentiation study using the relevant cell-based assays (cell surface binding, ADCC assay, receptor number determination and internalization assay) to discover the competitive advantage of their asset molecule
-Evaluate the outcome of biological assays and propose the strategies for improvement of their top candidate molecule
-Present the results delivered within the practical and a prospective research proposal to a broader audience (supervisors/peer scientists/investors)
Noch mehr Informationen zur Lehrveranstaltung, wie Termine oder Informationen zu Prüfungen, usw. finden Sie auf der Lehrveranstaltungsseite in BOKUonline.