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Dr Jenny Ottosson
Maria Berling (technician), Anna Bäckström (research engineer), Maria Carlsson (research engineer), Martin Hjelmare (technician), Hero Nikdin (technician), Katarina Pettersson (research engineer), Marie Skogs (technician), Johanna Steen (PhD-student), Hanna Tegel (PhD-student), Ulla Wrethagen (technician), Lan Lan Xu (technician), Louise Yderland (research engineer) and Feifan Yu (technician).
To express and purify PrEST-proteins to use for the preparation of antigens and affinity columns.
The Protein Factory group is responsible for protein expression and purification in the Human Proteome Resource program. Proteins are expressed as His6ABP fusions in E. coli shake flask cultures upon induction with IPTG. 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 hexahistidine affinity tag and metal affinity chromatography (IMAC) and performed under denaturing conditions. After 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.
Abbreviations:
ABP - Albumin Binding Protein
IPTG - Isopropyl-B-D-Thiogalactopyranoside
IMAC - Immobilized Metal Affinity Chromatography
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