Course: Bioanalysis, Practical (1502-462)

Persons:

• Prof. Dr. Lutz Fischer (verantwortlich)
• Dr. Sabine Lutz-Wahl (begleitend)
• Dr. Jens Pfannstiel (begleitend)
• Eva Proß (begleitend)

Type of Course:

exercise

In-Class Hours Per Week:

3

Contents:

The practical comprises the following bioanalytical experiments which are carried out, analysed, and documented independently by the students:
- Cultivation of Yarrowia lipolytica and sample preparation
- 2D gel electrophoresis for separation of proteins and protein isoforms.
- Tryptic digestion of proteins in gel slices
Separation of peptides by HPLC
- Maldi-ToF and ESI mass spectrometry combined with data bank analysis (e.g. MASCOT) for identification of proteins

The students understand the theoretical background and gain practical experiences in carrying out bioanalytical methods.
This includes:
- Using 1D- and 2D-electrophoretical methods for protein separation.
- Learning the principals and gaining practical experiences in immunological protein detection methods.
- Learning the principals and gaining practical experiences in MALDI-ToF mass spectrometry.
- Using MALDI-ToF mass spectrometry for protein identification and characterization of protein modifications.
- Learning the principals and gaining practical experiences in nano-HPLC coupled electrospray mass spectrometry.
- Learning how to perform database searching for mass spectrometry based protein identification.
- Using electrospray mass spectrometry for the absolute quantification of small molecules in extracts of biological fluids.

Literature:

Lottspeich, F., Zorbas, H.: Bioanalytik,: 3. Auflage (2012), Springer/Spektrum, München, ISBN-13: 978-3827429421

Location:

Hohenheim

Module:

• 1502-460 Bioanalysis (compulsory)