An Improved Protein Bioreactor EFFICIENT PRODUCT ISOLATION DURING IN VITRO PROTEIN BIOSYNTHESIS VIA AFFINITY TAG*

In vitro protein biosynthesis became a powerful technol-ogy for biochemical research. Beside the determination of structure and function in vitro selection of proteins is also of great interest. In most cases the use of a synthesized protein for further applications depends on its purity. For this purpose the in vitro production and purification of proteins with short affinity tails was established. A cell-free protein synthesis system was employed to produce bovine heart fatty acid-binding protein and bacterial chloramphenicol acetyltransferase with and without fu-sion of the Strep-tag affinity peptide. The quantitative removal of fusion protein during cell-free synthesis from a batch reaction and a semicontinuous flow cell-free reac-tor were achieved. No significant influence of the Strep-tag and the conditions during the affinity chromatograph