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August 22, 2016

Simplify chromatography method creation

By Tuomo Frigård, Scientist at GE Healthcare

Efficient method creation is often about finding useful tips on how to develop methods easy to maintain, reuse, and share. In this post, you can read about tips and tricks to simplify method creation as well as useful system functions that might have been forgotten.

Tips for ÄKTA systems installed with UNICORN software

The following tips make method creation more efficient and methods easier to use long-term.

  • In Method creation, you will find help for individual instructions by pressing F1.
  • Prepared method phases and blocks can easily be shared with others through Methods and Method libraries.
  • Use and update the column list to get relevant predefined column data quickly into your method.
  • Use Column Logbook for column tracking to ensure column maintenance is done based on performance.
  • Use a scouting method to scout for different parameter settings such as flow rate or buffer conditions.
  • Variables of scouting methods created in UNICORN 7 can later be used in evaluations to sort and compare results.
  • Use design of experiments (DoE) functionality to increase process performance and process knowledge by direct access to tools for setting up a study, performing runs, and evaluate results.
  • To minimize loss of sample remaining in the flow path, add an extra buffer volume to sample injection in the method block Finalize sample injection. Using an extra buffer volume when injecting sample onto the column is preferably used in bind-elute mode to avoid diluting the target molecule.

Hardware components are available for installation to support automated methods in the ÄKTA avant and ÄKTA pure system. Below are a few of my favorites:

  • The cooled fraction collector of ÄKTA avant enables samples to be left in the system at conditions beneficial for protein stability until further use.
  • The I/O box can be used to control external equipment such as a light scattering detector, autosampler, or a scale to apply sample directly from a perfusion bioreactor run.
  • Additional valves can be used in multiple combinations to increase process flexibility.
  • Loop valve to increase loop positions. If large volumes are required, the loops can be replaced by sample /buffer containers, for example, when doing automated multistep purifications or continuous processing.
  • Versatile valves can be used to create flow paths for more advanced setups such as automated multistep chromatography or to handle external in-line or off-line detectors.
  • Additional inlet valves can be used to generate extra column positions.

Chromatography requires training and education, but don’t worry. There is help available. Join ÄKTA club to participate in discussions around ÄKTA system and protein purification. GE Healthcare also offers basic and advanced trainings via Fast Trak.