As the worldwide leading suppliers of custom protein services, Creative Biostructure is pleased to provide free protein purification technical resources to facilitate your daily research based on our advanced MagHelix™ Protein Purification services. Affinity chromatography is involved with more than 60% of all purification techniques as stated, and affinity chromatography basically depends on a molecule bound to a column to recognize the target molecule.
There are three main steps in affinity chromatography purification:
Figure 1. Illustration of three main steps in affinity chromatography
The ligand that is used for purification of its respective binding partner can be any component. There are some common biological interactions can be frequently applied in affinity chromatography, as follows:
Figure 2. Illustration of a variety of molecules that can be purified by using affinity chromatography. (Affinity Chromatography Principles and Methods, 18-1022-29)
Affinity chromatography in practice
Figure 3. Illustration of a typical affinity purification. (Affinity Chromatography Principles and Methods, 18-1022-29)
Figure 4. Illustration of simple procedures to perform affinity purification on a pre-packed HiTrap column. (Affinity Chromatography Principles and Methods, 18-1022-29)
Things to Consider:
Troubleshooting:
(Data were taken from Affinity Chromatography Principles and Methods, 18-1022-29)
1. Elution with a sharp target peak--Satisfactory result
2. Elution with a broad, low target peak while appling a binding buffer
3. Elution with a broad, low target peak
4. Elution with a broad, low target peak while still under binding conditions
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