School of Biotechnology
Proteomics
Research

  Protein Factory
Summary
Administration
Al-Khalili Szigyarto
Berglund
Hober
Lundberg
Nilsson
Odeberg
Ottosson
Persson
Schwenk
Uhlén

Principal Investigator (PI): Dr. Jenny Ottosson

Personnel: Johanna Steen (PhD-student), Hanna Tegel (PhD-student), Anna Konrad (research engineer), Asif Halimi (technician) and Ulla Wrethagen (technician)

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

Description: 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 electrospray 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

Last updated: 2010-12-22