Equipment for preparative purification of tagged and untagged proteins, including the ÄKTA series of liquid chromatography systems from Cytiva Life Sciences, are frequently used in settings ranging from scientific research laboratories to production plants. With these systems, it is possible to perform a variety of chromatographic separation modes including affinity, size exclusion, gel filtration, hydrophobic interaction, reversed phase, and ion exchange. However, in all modes and setups, the formation of bubbles may hamper the reproducibility of purification results and thus limit the overall productivity.
Bubbles form because of high levels of dissolved gases in the employed buffers and solvents. The multiple pressure drops occurring within the liquid flow path through a chromatography system (injector valves, separation column, detector cell, etc.), may initiate bubble formation, especially if any segment also has a higher temperature compared to the eluent bottle. Traditional ways of removing dissolved gases have been to treat the buffers, eluents and mobile phases in ultrasonic baths, but this is both labour intensive and rather inefficient, and risk changing the solution temperature, or may even accelerate degradation of sensitive components. The most efficient and user-friendly way to remove bubbles and dissolved gases in liquid chromatography is by using online vacuum degassers.
The DEGASi® range of online vacuum degassers from Biotech Fluidics can be fitted to any brand of liquid chromatography instruments to solve all your troubles with bubbles. The DEGASi® PREP stand-alone degasser is designed for flow rates up to about 20 ml/min per channel and thus suitable for most of the laboratory protein purification chromatography systems including ÄKTA start, ÄKTA go, ÄKTA pure 25 and ÄKTA avant 25, plus discontinued legacy systems such as ÄKTAprime plus, ÄKTAxpress ÄKTA Purifyer and ÄKTA FPLC (some these models were built by GE Healthcare or Amersham Pharmacia Biotech). Several alternative DEGASi® degasser designs are available for chromatography systems with higher or lower flow rate specifications.