Protein Purification Lab Course

S. Hoffmann (Structural Biochemistry)
D. Willbold (Structural Biochemistry)

The course will cover overexpression of proteins, protein purification and characterisation. The course consists of a practical part combined with several lectures in which the theory behind the methods used is discussed.
 
The theoretical part covers the topics:

• Introduction to Protein Expression Systems
  
  • Bacterial Expression Systems (main focus)
  • Cell-free Production of Proteins
  • Insect-based Expression Systems
  • Mammalian Expression Systems
• Protein Purification Strategies
  
  • "salting out"
  • gel filtration
  • ion exchange
  • hydrophobic interaction
  • reversed phase chromatography
  • use of affinity tags
• Methods for sample characterisation
  
  • protein concentration
  • SDS gel electrophoresis
  • western blotting
  • spectroscopic methods

Laboratory work will be performed on a protein, which will be recombinantly expressed in E. coli. The recombinant protein will be purified from a crude form (cell lysate) to near homogeneity using different purification strategies and will subsequently be characterised in respect to yield and purity. The course is a 5 full-day course and takes place in Jülich.

February 7-11, 2011