EXPORT-ORIENTED MANAGEMENT - IMC Fachhochschule Krems

Process online monitoring: Relevant on-line probes and their usage for process controll
get to be known, the usage of on-line measurements instead of off-line measurements -
how to replace them
Prerequisites/ Level
Advanced
Course Description
Fermentor instrumentation / - set up; Definition in-, on-, at- and off-line; Sampling
techniques; Off-line analytics, Chromatographic methods (substrate, nucleotides,…);
-Omics (Transcriptomics (microarrays, real time PCR,…), proteomics, metabolomics); in-
and on-line methods; spectroscopic methods; software sensors; multivariant data
analysis / chemometrics; FDA´s QbD and PAT guideline and initiative
Teaching Concept
Academic lectures, problem sessions, group works
Evaluation
Group works, final exam
Recombinant protein production and modification laboratory
Semester Hours per week Subject code ECTS
Fall 4 PROT_L 5
Learning Outcomes/ Course Objectives
In the laboratory course the students will get familiar with the protein expression
platform P. pastoris. The human cytokine TNF alpha will be produced intracellular and the
product will be purified via affinity purification. Finally the product will be characterized in
a cell based test system (WEHI cells). This murine cell line is very sensitive for apoptosis
induction via human TNF alpha (IC50 value is ~10pg).
Prerequisites/ Level
Advanced
Course Description
Transformation (chemical) of Pichia
Media Design, Cultivation of Pichia
Screening of Expression clones
Induction of recombinant protein production
Disruption of cells via French Press or glass beads
Affinity Purification of TNF alpha
Buffer exchange via gel filtration
Activity testing via cytotoxicity assay in murine cell line (cell culture technique)
Teaching Concept
The students will be prepared for the laboratory course in the corresponding lecture
Evaluation
Protocol and review of provided papers
FHR-5-0008_Vers.03_Rev.00_2012 547