Purification and Characterization of Recombinant Proteins

We are organizing (again) a practical hands-on protein purification course from December 4th to 20th. We maximally can accommodate 16 participants, which will form groups of 2 to 4 participants. Each group needs 3 full days to go thru the practical exercises, but day 3 of the course will be overlapping with day 1 of the next group. Venue is Biomedicum Helsinki, rooms A516a1 (where the machinery is) and B318a/b (our lab). We will purify a protein (VEGF receptor 3) using a two-step protocol (affinity chromatography + gel filtration) on the the Äkta Avant FPLC device. On the third course day we’ll assay its interaction with its ligand (VEGF-C) on the ITC (isothermal calorimetry) device. We have given a similar course in 2015 ( http://www.helisci.fi/hbgs/FPLC2015/)  . This new course ( https://courses.helsinki.fi/en/DPBM-135/120171139  ) will have a similar structure, but we expand it by one day to analyze protein interactions making use of the new isothermal calorimetry device of our B3P core facility. The documentation and results will be also available from here . Since we can run maximally two samples at a time (we have “only” two FPLC machines), we will have to split the participants into groups (of 2-4 students/group) and repeat the 3-day course several times depending on the number of participants.

Day 1 (3 hours) 14:00 - Preparation of the FPLC machine 16:00 - Start of the affinity chromatography column sample loading

Day 2 (6 hours) 8:00 – 9:00 - Affinity chromatography column washing and elution 9:00 – 11:00 - Size exclusion chromatography 11:00 – 16:00 - SDS-PAGE & Coomassie staining

Day 3 (5 hours) 9:00 – 12:00 - ITC run using the purified proteins (antibody – antigen interaction or receptor – ligand interaction) 13:00 – 15:00 – Discussion and analysis of results