Principal Investigator (PI): Dr. Jenny Ottosson

Personnel: Johanna Steen (research engineer), Hanna Tegel (research engineer), Maria Stenvall (research engineer), Robin Eriksson (technician), Ulla Vrethagen (technician).

Responsibility: To express and purify PrEST-proteins in fusion with His₆ABP to use for the preparation of antigens and affinity columns.

Description: The protein factory unit is responsible for protein expression and purification in the Human Proteome Resource (HPR) facility. The proteins are expressed in E. coli in batches of shakeflask cultures (100ml). A fully automated protein purification system has been developed to allow for purifications of up to 60 cell lysates at a time. One-step purification is enabled by the histidine-tag and metal affinity chromatography (IMAC) and performed under denaturing conditions. After an evaluation of protein concentration, purity, and solubility the molecular weight of the PrEST proteins are determined by mass spectrometry as a final quality control. The purified proteins are then used to prepare antigens and affinity columns with PrEST-ligands. In addition Protein Factory produces affinity resin with His₆ABP-ligand..