Pilot Scale Experiments To Improve Chromatography Column Packing Procedures

Large scale liquid chromatography is the dominant purification technique used in the production of recombinant proteins. The quality of the column packing is a key parameter and its integrity impacts the purification performance. A ten cm diameter Index column (GE Healthcare) was packed with three different types of resin and three packing methods were executed for each resin. After each pack, the column was integrity tested by the Height Equivalent of Theoretical Plate method (HETP).  The integrity test results showed that in most cases, packing by axial compression resulted in an  increased number of theoretical plates. HETP integrity testing was also performed at varied linear flow rates for each column pack. A decreased flow rate was correlated with increased number of theoretical plates. Consistent methodology using the same residence time on the column is recommended for comparison of column integrity test results with similar column scales (diameter and bed heights) between  projects.