Continuous separation of green fluorescent protein by annular chromatography

The concept of annular chromatography was tested by separation of a real protein solution used in biotechnology. Green fluorescent protein was expressed in S. cerevisiae and the extract was continuously separated by a pressurized annular chromatograph packed with a Superdex 200 prep grade size-exclusion chromatography medium. Purity was checked by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, Western blotting and fluorescent intensity. The continuous mode was compared to batchwise operation. Under the assumption that equivalent fractions are collected, both modes are comparable.