Proteome of BCG-derived membrane vesicles

Proteins in two different BCG-derived membrane vesicles, extracellular vesicles, were compared. One is isolated from statically cultured BCG in Sauton's medium (Sample #1 and #2), commonly used to produce BCG vaccine. Another is isolated from planktonically cultured BCG in Muller-Hinton II Broth (Sample #3 and #4), commonly used for bacterial experiments. Proteins from BCG-MVs were separated via SDS-PAGE on a 12% polyacrylamide gel (10 µg in each lane). The resulting gel was stained with 10% Coomassie Brilliant Blue (G-250), and 4 pieces per lane were excised. The samples were reduced, alkylated and trypsinized via the in-gel digestion method. The peptides extracted from the gel were subjected to nanoLC-MS/MS analysis with a system consisting of an Orbitrap Velos Pro (Thermo) coupled with an Advance UHPLC (Brucker).