Department of Natural Sciences
Medical Proteomics (MSc)
Lecture
Compulsory Course
- in MSc Biomedical Sciences, 3rd Semester
- 6h/week (2L/2E/2P)
- Credits: 8 ECTS
Learning outcomes:
Successful students can
- find suitable research articles in the literature and select the necessary information in publications for state of the art sample preparation for electrophoretic, chromatographic and mass spectrometric analyses
- write detailed lab scripts for sample preparation based on published procedures in the literature
- design reasonable proteomics work-flows for selected introductionary applications in the field of mining and differential proteomics
- decide on reasonable methods for protein purification
- explain typical current proteomics strategies and applications to a novice in the field
Content:
- Sample preparation: sampling, sample handling, cell lysis methods, laser microdissection, diverse prefractionation techniques (like i.e. affinity and gel filtration chromatography, protein depletion, ultracentrifugation), protein digestion, protein quantification, clinical requirements
- Separation techniques: 1- and 2D gel electrophoresis, DIGE, preparative IEF, capillary electrophoresis, HPLC and UPLC (employing Reversed Phase and Ion Exchange), multidimensional LC
- Mass spectrometry: general principles, MALDI and ESI ionization, TOF, quadrupole, FT-ICR/ orbitrap, ion trap, tandem and hybrid instrumentation, CID, ECD/ETD, LC-ESI, SELDI, MALDI imaging
- Proteomics strategies: bottom up, middle down, top down, discovery vs. hypothesis based, identification based on PMF and peptide sequencing, label and label free quantification approaches like SRM/MRM, SILAC, iTRAQ, ICAT, database analysis, search algorithm
- Applications: mining, differential like biomarker discovery, protein-protein interactions, posttranslational modifications
Internship
Lab Class
Content:
- Isolation and purification of Benzoylformate decarboxylase from P. putida, overexpressed in E. coli. Determination of the purity, activity and overall yield.
- 2D-gel electrophoresis analysis of E. coli cell lysates, analyzing differences of the bacterial proteome in response to overexpression induction in cell culture
Requirements
none
Recommendations:
- Basic Organic Chemistry
- Instrumental Analysis
- Clinical Chemistry
- Biochemistry
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The grade of the final exam determines the overall grade of the course
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A pass of six on-line tests and a lab report is required
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Attendance of lab class is mandatory
Literature
- Lovric, J. (2011): Introducing Proteomics from Concepts to Sample Preparation, Mass Spectrometry and Data Analysis, Wiley-Blackwell
- Westermeier, R. (2016): Electrophoresis in Practice: A guide to Methods and Applications of DNA and Protein Separations, Wiley-VCH
- Greaves, J., Roboz, J. (2014): Mass Spectrometry for the Novice, CRC Press
Links
Further links