1 GE Healthcare Bio-Sciences AB, Björkgatan 30, SE-751 84 Uppsala, Sweden
2 Dr. von Haunersches Kinderspital, 80337 Munchen, Germany
3University of Leiden, NL 2333CC Leiden, The Netherlands
Sponsored by GE Healthcare Published in July 2011 (p.17) Sponsored,vendor-submitted protocol
In this work, MBP-tagged proteins expressed in E. coli were purified on prepacked 1 ml or 5 ml MBPTrap HP columns. A stability study was performed with six repeated purification runs on the same column, each followed by regeneration with 0.5 M NaOH. The results showed no significant change in yield or purity.
A purification of medium-chain acyl-CoA dehydrogenase (MCAD), a protein involved in metabolic disease, was performed. The total purification time was reduced due to elution of highly concentrated target protein, thereby eliminating the need for a concentration step and simplifying the whole purification procedure. Furthermore, an automated two step purification of apoptin was performed on ÄKTAxpress™ where the affinity chromatography step on MBPTrap HP was followed by a gel filtration step.
In summary, the results show that prepacked MBPTrap HP columns decrease purification time and give high purity and reproducibility.