BMB545: Strategies for Protein Purification and Characterization
The Study Board for Science
Teaching language: Danish, but English if international students are enrolled
EKA: N200034112, N200034102
Assessment: Second examiner: None, Second examiner: External
Grading: Pass/Fail
Offered in: Odense
Offered in: Autumn
Level: Bachelor
STADS ID (UVA): N200034101
ECTS value: 10
Date of Approval: 13-05-2020
Duration: 1 semester
Version: Archive
Comment
Attention! For Autumn 2024 the participant limit is 6 participants
The course has limited entry. The following criterias are taken into consideration when seats are assigned.
- Students with the most ECTS from their bachelor at the beginning of the semester
If students are equal – seats are allocated based on random draw.
The academic envirometns at The faculty of science manages the prioritisation and at waiting list is established and will then be made aware from the faculty. The waiting list will not be transferred to the following year.
Max 10 and minimum 8 people pr. lab pr. week. (approx. 60 hours of exercise per week).
Co-read with BMB841.
Entry requirements
Academic preconditions
Students taking the course are expected to:
- 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.
Participant limit
Course introduction
The aim of the course is to enable the student to understand the theory and use of classical protein purification methods and to apply them in practice, which is important in regard to understanding and characterization of protein structure and functions and the development of e.g. protein based drugs.
The course builds on the knowledge acquired in the courses KE501 or similar courses with a focus on general and organic chemistry, BMB532 or similar courses on basic biochemistry and BMB536.
The course builds on knowledge of protein chemistry and bioinformatics achieved in courses such as BMB533 and BMB511, and gives the academic basis for subsequent electives within the field (i.e BMB836) as well as a masters project.
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
The learning objectives of the course is that the student demonstrates the ability to:
- 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.
Content
The following main topics are contained in the course:
- 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.
Literature
- Robert K, Scopes: Protein Purification - Principles and Practice, Springer Verlag, 3. udgave.
- Udvalgte forsknings- og oversigtsartikler.
See Itslearning for syllabus lists and additional literature references.
Examination regulations
Prerequisites for participating in the exam a)
Timing
Autumn
Tests
Participation in practical exercises
EKA
N200034112
Assessment
Second examiner: None
Grading
Pass/Fail
Identification
Full name and SDU username
Language
Normally, the same as teaching language
Examination aids
To be announced during the course
ECTS value
0
Additional information
The prerequisite examination is a prerequisite for participation in exam element a)
Exam element a)
Timing
Autumn
Prerequisites
| Type | Prerequisite name | Prerequisite course |
|---|---|---|
| Prerequisite not found |
Tests
Oral examination based on the lab report
EKA
N200034102
Assessment
Second examiner: External
Grading
Pass/Fail
Identification
Full name and SDU username
Language
Normally, the same as teaching language
Examination aids
To be announced during the course
ECTS value
10
Additional information
The oral exam is based on the report and it's not possible to pass the course without
Indicative number of lessons
Teaching Method
Activities during the study phase:
- Self-study of the textbook
- Written report
- Individual recap of introductory and training phase
- Repetition up for the exam