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GE Healthcare Life Sciences Part of GE Healthcare
labsep
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
Animation
Basic Principles
The Separation Mechanism
Type of ion exchangers
Elution modes
The typical Ion exchange experiment
Charge properties of proteins and peptides
Effect of running pH
Resolution in IEX
Optimisation of IEX experiments
Ion Exchange in practice
Technique Profile
What is Ion Exchange?
Reversed phase chromatography
Protein Purifier software
BioProcess™ Glossary
The typical Ion exchange experiment

IEX is the most frequently used in gradient mode and the experiment consists of four phases as shown below.

Explanation of symbols

Symbolic representation of a section of an anion exchanger bead.
Sample component with net charge = -1.
Sample component with net charge = -2.
Sample component with net charge = -3.
Positively charged
or neutral sample components.



1. Equilibration
Buffer A is pumped through the column until pH and salt conc. is correct.		2. Sample application and wash
Sample is added and adsorbed.
3. Gradient elution
Gradient is started and adsorbed sample components are eluted in order of their respective net charges.
Elution order: 		4. Regeneration
Charged components still remaining are washed out.
Fig 4.1. The separation of four proteins with different negative net charges using gradient mode.