940331 Cell factory - plants


Type
Exercise course
Semester hours
3
Lecturer (assistant)
Organisation
Offered in
Sommersemester 2023
Languages of instruction
Englisch

Content

The COVID-19 pandemic has shown how important it is for biotechnological research / industry to act quickly to contain it. A key factor is the efficient production of SARS-CoV2 relevant proteins for both diagnostic (e.g. virus antigens) and therapeutic (protective antibodies) purposes. This course refers to this development and introduces a plant-based expression system to efficiently produce SARS-CoV2 relevant proteins for diagnostic purposes.
Two classes of proteins are produced: (i) a viral spike protein domain (SP), which plays a key role in viral infection, and (ii) a specific antibody that binds to this domain. The possible use of the two components for diagnostic purposes is then shown (ELISA binding studies).
Correct subcellular localization plays an important role in the expression of recombinant proteins, as this can significantly influence the properties of proteins. With reference to this, the subcellular transport and deposition of various recombinantly produced proteins are examined during the course.
Specifically, the following experiments are carried out:
1) Plant-based expression and purification of a SARS-CoV2 specific antibody and a SARS-CoV2 SP domain
3) ELISA binding studies to develop a diagnostic method
2) Plant-based expression of fluorescent reporter proteins (e.g. fusion of SP with a fluorescent protein) with various subcellular targeting sequences with subsequent detection of the localization using confocal microscopy.

The practical experiments are accompanied by a detailed theoretical part that presents the background of the plant expression system and the production of important biomolecules in times of pandemics

Previous knowledge expected

Basic knowledge about cell and molecular biology;
practical experience in labwork

Objective (expected results of study and acquired competences)

Participants
•recognize the potential of plants as a platform for the rapid production of recombinant proteins
•are familiar with the use of different protein classes in diagnostics (and therapy)
•understand the principals of ELISA test kits
•know methods for the visualization of proteins
•understand the basics of protein purification (immuno-affinity methods) and quantification
•are familiar with the visual representation of subcellular compartments (such as Golgi apparatus, ER, Apoplast)
•recognize the potential of fluorescent fusion proteins to investigate cell biological processes (such as subcellular protein transport)
You can find more details like the schedule or information about exams on the course-page in BOKUonline.