BMB202: Protein Purification
The course has limited entry. The following criterias are taken into consideration when seats are assigned.
- PhD students after time of enrollment
- Students with the most ECTS from their master
- Students who follows master courses concurrent with their bachelor programme (dispensation to 30 ECTS Master courses)
- be familiar with basic general and organic chemistry and biochemistry in particular fatty acid metabolism, organic molecular structures and functional groups and pH concept.
- be familiar with the importance of organic molecules in biochemistry including biological macromolecules.
- be familiar with simple inorganic molecules and salts as well as their properties.
- have practical experience with laboratory work and general laboratory safety.
The course builds on the knowledge acquired in the courses FF503 or similar courses with a focus on general and organic chemistry, BMB532 or similar courses on basic biochemistry and BMB505/536.
In relation to the competence profile of the degree it is the explicit focus of the course to:
- obtain skills to apply advanced methods to purify and characterize proteins
- provide skills in creating the experimental basis for the practicality of purification of proteins
- gain knowledge about the techniques of protein purification and characterization of their properties including enzyme activity.
- to understand and on a scientific basis reflect on biochemical and molecular biological knowledge, and to identify scientific problems.
Expected learning outcome
- Discuss and analyze how fatty acid synthesis takes place in bacteria, simple eukaryotic cells like yeast cells, and mammalian cells including various fatty acid synthesis enzyme systems, respectively, in both the cytosol and mitochondria.
- Discuss the mechanisms behind fatty acid synthesis, elongation and termination
- Select precise methods for homogenization of specific tissues and cells
- Evaluate the advantages and disadvantages of ion exchange, gel filtration and affinity chromatography
- Discuss and analyze the theory behind column chromatography and factors affecting purification and separation of proteins.
- Discuss the theory of protein solubility and how this may be affected by salt, organic solvents, temperature, and pH.
- Discuss and analyze the principles of the electrophoretic separation of molecules, including proteins.
- Discuss and put into perspective the use of electrophoresis in the understanding of protein properties and structure.
- Discuss and assess typical techniques used for determining the concentration of proteins.
- A theoretical part where the theory behind fatty acid synthesis is reviewed.
- The applied methods used are reviewed.
- A practical part where a particular protein is purified using the methods above. The methods used include fractionation of cellular components by centrifugation, and protein fractionation by precipitation, ion exchange chromatography and gel filtration chromatography. Furthermore, participants will get to perform photometric determinations of protein and enzyme activities. The purity of the purified protein and its molecular weight is determined by gel electrophoresis.
- Robert K, Scopes: Protein Purification - Principles and Practice, Springer Verlag, 3. udgave.
- Udvalgte forsknings- og oversigtsartikler.
See Blackboard for syllabus lists and additional literature references.
Prerequisites for participating in the exam a)
Participation in practical exercises
Prerequisites for participating in the exam b)
Submission of written report
Exam element a)
|Type||Prerequisite name||Prerequisite course|
|Examination part||Prerequisites for participating in the exam b)||N220000101, BMB202: Protein Purification|
Oral examination based on the lab report
Reexam in the same exam period or immediately thereafter.
The examination form for re-examination may be different from the exam form at the regular exam.
Indicative number of lessons
- Self-study of the textbook
- Written report
- Individual recap of introductory and training phase
- Repetition up for the exam