Programme Lille

 

Modern large-scale proteomic approaches offer a unique opportunity to access the protein content and regulation in physiological and pathophysiological context. Nowadays, the identification and relative quantification of several thousands of proteins can be obtained in a single automated acquisition run of max 2H. These strategies rely on the use of mass spectrometry coupled to hyphenated techniques, and liquid chromatography coupled to mass spectrometry (LC-MS) remains so far, the state-of-the-art strategy. To achieve large-scale identification, shot gun approaches are conventionally privileged. In shot gun after the proteins are extracted, these are submitted to reduction and alkylation followed by global enzymatic digestion (generally trypsin). The digested peptides are then analyzed by LC-MS and the MS data further submitted to databank interrogation for identification. The workshop will offer to the participants to perform themselves the complete experimental workflow from sample preparation to data processing using iST for the sample processing.
The attendees are offered to work with their own samples. However, because of the limited time, attendees’ samples will be processed prior to the workshop and model samples will be prepared during the workshop except for the data processing.

 

PROGRAMME

5 July 2022 | Tuesday
09:00-09:30       Welcome coffee
09:30-10:00       Objectives and schedule of the training
10:00-11:00       Review of the trainee samples and presentation of the technical workflow
11:00-12:00       Lab experiments: Spatial Digestion CHIP1000
12:00-13:30       Lunch Break
13:30-14:30       Tissue extraction
14:30-17:30       FASP (reduction/alkylation, washing, digestion) Sample kept overnight
                           for digestion + microextraction with LMJ
17:30-18:00       Debriefing

 

6 July 2022 | Wednesday
09:00-09:30       Welcome coffee and objective of the day
09:30-12:00       Last steps of sample preparation, Zip-Tip, Loading of samples on the LC-MS
12:00-13:30       Lunch Break
13:30-17:30       Data processing learning (MaxQuant / Perseus) + processing of trainee’s data
17:30-18:00       Debriefing
19:00-22:00       Walk through the “Heron Park” and Dinner

 

7 July 2022 | Thursday
09:00-09:30       Welcome coffee and objective of the day
09:30-12:00       Analyzing the data of the sample acquired during the workshop
12:30-13:30       Lunch Break
13:30-14:00       Training conclusion