Purification and Characterization of Recombinant Proteins

Size exclusion chromatogram (acetone performance test)

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 course 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.

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


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