GE Healthcare
 
GE Healthcare Life Sciences Part of GE Healthcare
Education Centre
About the purification of biomolecules
Purpose of purification
Developing purification protocols
How to combine purification steps
Purification development - summary
LC techniques
Affinity Chromatography
Desalting & Gel Filtration
Hydrophobic interaction chromatography
Ion exchange chromatography
Reversed phase chromatography
Protein Purifier software
Tutorial

Purification development - summary

In the Purification centre you will find complete purification protocols for different situations.

However, when the only option is to develop a new protocol “from scratch” the tables below can serve as a checklist.

Before you start
  • Develop assay methods
  • Set the aims (purity and quantity)
  • Characterize the target protein
  • Determine the sample stability window

Protocol development checklist
  • Select techniques and conditions compatible with sample stability.
  • Remove proteases at an early stage or yield may suffer.
  • Reduce volume in early steps.
  • Use combinations of different separation principles.
  • Start with high selectivity – increase efficiency.
  • Use few steps.
  • Limit sample handling between purification steps.