Figure 2 shows the method sequence between both the filtration and chromatography systems. Each method is numbered to the left of each box to illustrate the order in which it was executed.
The preproduct equilibration of the filter automatically started the water flush of the ÄKTApilot system from storage, followed by equilibration of the chromatography column.
The product section began with clarification/diafiltration of the yeast harvest with passage of the target protein into the permeate. An intermediate vessel between the two systems contained the permeate outlet as well as the chromatography inlet tubing. Protein was pumped onto the column from the intermediate vessel until detection of air terminated the load.
The postproduct sequence began with a water flush of the hollow-fiber filter. This method was started via Ready signal from the purification method. It was followed by CIP of the filter. At this time the purification method was completed and a signal was sent to begin CIP of the chromatography column.
CIP of the filter was completed first, initiating filter storage. At this time CIP of the column was completed. The ÄKTApilot system received a signal from the filter storage method to begin storage of the system and column.
Some manual interaction was required to move the inlet line on UniFlux 10 to the next buffer or sample, and the operator was alerted via an on-screen message. The current UniFlux platform offers a standard option of a four-valve inlet panel for increased automation. The ÄKTApilot system only required manual interaction at the start of the equilibration method and for the inlets to storage method at the end of the process. Again, the operator was notified via on-screen messaging. The method was paused until the operator confirmed the correct configuration.