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I5: Introduction to Protein Interaction Analysis by Affinity Purification – Mass Spectrometry – 34604

(3 days; 1 CP; Spring 2023)
Course dates: 18.4. - 20.4.2023

Thomas Bock, Alexander Schmidt

3-day practical course on affinity purification-mass spectrometry (AP-MS). During the course we will look into the methodology used for identification of protein-protein interactions by mass spectrometry. Specifically, we will perform the purification of a target protein and its preparation for mass spectrometry, we will perform the mass spectrometry measurement of the sample, as well as learn how to analyze the resulting AP-MS data.

Day 1: Sample preparation
Seminar: Introduction to AP-MS workflows, advantages/disadvantages of different protein affinity tags/enrichment strategies

Hands-on training: Affinity purification of a target protein out of a cellular lysate, protein digestion over night

Day 2: Mass spectrometry analysis
Seminar: Introduction to mass spectrometry (MS) and its applications in molecular biology

Hands-on training: Solid-phase extraction of peptides, mass spectrometry measurement

Day 3: Data analysis
Seminar: Introduction to MS-data analysis, principles of protein identification, protein-protein interaction data analysis