Protein expression

The Protein Expression laboratory contains all the facilities needed to produce heterologous proteins from E. Coli or appropriately engineered bacteria through the common molecular biology techniques. The growth of bacteria is carried out at controlled temperature and under suitable stirring conditions (MaxQ 6000, Thermo Scientific). The isolation of the bacteria from the relative cultures, as well as the removal of the cellular debris after the bacteria lysis, is carried out using refrigerated centrifuges for large (SL-40R, Thermo Scientific) and small (MicroCL-21R, Thermo Scientific) volumes. The over-expressed proteins can be purified by several kind of chromatographic separations (SEC and ion-exchange columns with different loading and resolution capacities) by means of an automated FPLC purification system suitably refrigerated to 4 °C.

Fast Protein Liquid Chromatography, Biologic Duo Flow

The system is used for a fast purification and characterization of a protein mixture. The pumps can go up to 3500 psi with a flow rate of 10 ml/min. The instrument includes a conductivity meter as well as a UV detector along with a fraction collector, all controlled by a common software platform. Small (0.1 ml) to large (25 ml) samples can be directly loaded. DuoFlow system is stored at 4°C, thus allowing the correct conditions of the protein purification. The large set of columns can be used to perform ionic separations, size-exclusion and affinity chromatography.