Protein Purification and Characterisation

Course Description

COURSE CONTENT

Lectures: Isolation and solubilisation of proteins from natural materials. Separation of proteins by precipitation techniques. Separation of proteins by centrifugation and by using semipermeable membranes. Separation of proteins by chromatographic methods. Genetic tagging of proteins for purification. Electrophoretic methods. Application of analytical methods for protein characterization (NMR, MS, CD spectroscopy, X-ray crystallography). Designing of protein purification procedures.

Seminars: The seminars are focused on solving problems in the field of application of various preparative and analytical methods for protein separation and purification (precipitation methods, membrane separation, chromatographic methods, electrophoretic methods). Computer simulation of the protein purification process.

Practical courses: separation and purification of proteins from egg white to homogeneity using precipitation and chromatographic methods, centrifugation and semipermeable membranes. Electrophoretic analysis and monitoring of the purification process, analysis of purification efficiency after each step of purification.

LEARNING OUTCOMES

• propose the protein purification procedure starting from natural materials or cultivated biomass
• select and conduct common preparative methods for protein separation; identify advantages, limitations and capabilities of each method
• select and conduct common analytical methods for protein separation in order to monitor the progress of purification and apply appropriate analytical methods for characterization of purified proteins
• evaluate efficiency of the purification procedure and modify it in order to increase efficiency of the method, yield or quality of the final product and decrease the cost of the purification procedure

To enrol in this course, the following courses must be completed:

• Biochemistry 1

Literature

1. Guide to protein purification (Deutscher M.P. ured.) Methods in Ezymology 182, Academic Press Inc., San Diego, 1990.

2. Basic Methods in Protein Purification and analysis (Simpson R.J., Adams P.D., Golemis E.A. ured.), CSH Press, New York, 2009

3. Short protocols in protein science (Coligan J.E., Dunn B.M., Speicher D.W., Wingfield P.T. ured.), Wiley, 2003

4. D.L. Nelson, M.M. Cox: Lehninger Principles of Biochemistry (4th edition), Worth Publisher, New York, 2005.

5. J.M. Berg, J.L. Tymoczko, G.J. Gatto, L. Stryer, Biochemistry (eighth edition), W.H. Freeman and Co., New York, 2015.