Three groups of properties of the target molecule are used in affinity chromatography:
1. Specific binding properties based on biological activity like:
- Enzyme active sites
- Receptor binding sites
- Antibody binding sites etc.
These are used together with the natural ligand or an analogue of it. Sometimes the analogue has a broader specificity and can be used for group separations.
2. Naturally occurring prosthetic groups like: polysaccharides etc.
Such properties normally allow group separations only.
3. Molecules equipped with an affinity tag like:
- Glutathione-S-Transferase (GST)
- Oligo histidine etc.
This group of properties is used almost exclusively for recombinant fusion proteins.
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Mono-specific separations |
- Antigens
- Antibodies
- Hormones
- Receptors
- Enzymes
- Tagged recombinants
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Group-specific separations |
- Glycoproteins
- IgG antibodies
- Enzymes
- Proteins/peptides with accessible histidines
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