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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
Animation of affinity chromatography
Basic principles of affinity chromatography
The affinity chromatography separation mechanism
The affinity chromatography experiment
Types of target molecular properties
Types of affinity chromatography ligands
Suitable conditions for binding/elution
Affinity chromatography applied to recombinant proteins
Affinity chromatography in Practice
Affinity chromatography technique profile
What is affinity chromatography?
Desalting & Gel Filtration
Hydrophobic interaction chromatography
Ion exchange chromatography
Reversed phase chromatography
Protein Purifier software
BioProcess™ Glossary

Types of affinity chromatography ligands

Ligands used for mono-specific affinity chromatography are structurally and biologically closely related to the target molecule to be purified. This makes the selection of the ligand specific for each case. This also makes it commercially difficult to produce affinity media for mono-specific separations. However, media ready for a variety of coupling chemistries (see table below) are commercially available for the benefit of the user, who intends to run mono-specific affinity chromatography.


With small ligands (<1,000) there is a risk for steric interference like binding between matrix and target molecule or low accessibility of the ligand. Introducing a spacer arm will minimize these risks (Fig 3.2).

Principle of preparing affinity chromatography media

Fig 3.1. The sequence of events used to prepare
affinity chromatography media. Pre-activated media are
commercially available.


Spacer arm principle

Fig 3.2. A spacer is sometimes used to
ensure full accessibility of the
affinity ligand.

Ligands used for group-specific affinity chromatography have a much wider applicability and affinity media for this purpose are consequently commercially available.
The table below lists examples of commonly used ligands of this type.