BMB202: Protein Purification (10 ECTS)

STADS: 01013901

Level
PhD course

Teaching period
The course is offered in the autumn semester.

Teacher responsible
Email: nils.f@bmb.sdu.dk

Timetable
Show entire timetable
Show personal time table for this course.

Comment:
Max 12 og min. 8 på hvert hold i lab. pr. uge (ca. 60 øvelsestimer pr. uge). Holdinddeling til laboratorieøvelserne finder sted ved forelæsningerne. Øvelserne afholdes i hele uge 43.

Prerequisites:
A Bachelor’s degree in Biochemistry and Molecular Biology, Biomedicine or similar.

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.

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 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
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.

Subject overview
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.

Website
This course uses e-learn (blackboard).

Prerequisites for participating in the exam

1. Participation in practical exercises. (01013912).
2. Submission of written report. (01013922).

Both a) and b) must be passed in order to take part in the examination. Pass/fail, internal marking by teacher.

Assessment and marking:

1. Oral examination based on the lab report. External examiner. Grades according to the Danish 7-point scale. (10 ECTS). (01013902).

Reexam in the same exam period or immediately thereafter.

Expected working hours
The teaching method is based on three phase model.
Intro phase: 10 hours
Skills training phase: 60 hours, hereof:
 - Laboratory exercises: 60 hours

Educational activities

Educational form
Activities during the study phase:

• Self-study of the textbook
• Written report
• Individual recap of introductory and training phase
• Repetition up for the exam

Language
This course is taught in English, if international students participate. Otherwise the course is taught in Danish.

Remarks
The course has limited entry. The following 4 criterias are taken into consideration when seats are assigned.

1. Students with the most ECTS from their master
2. Students who are accepted conditionally on the master 
3. Students who follows master courses concurrent with their bachelor programme
4. Bachelor students

If the score is even lots are drawn.

The academic enviroments of 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.

In order to keep the assigned seat you need to attend the first course day or notify the teacher, otherwise the seat is given to the next student from the waiting list.

Course enrollment
See deadline of enrolment.

Tuition fees for single courses
See fees for single courses.