Page 114 - Phytochemistry 2 (06-PG 605)

4- Affinity chromatography (LSC)
(Bioselective adsorption)

Based on the specific biochemical reactions between antigen &
antibodies, enzymes, substrates
Used for purification of enzymes, antigen (drug & metabolites) anti
bodies, hormones, viruses' parts, proteins, that are present in
trace amount
Principles:
1-Select stationary phase
St. ph. is a gel matrix to which a ligand is covalently bonded
through a spacer group.
*The ligand must have a specific binding affinity to the substrate
2- Sample is applied to the top of the column.
3- On development,compound with specific affinity to the ligand,
become bound and retained.
The unbound compounds will be washed through the column
4- The composition or PH of mobile phase is changed to weaken
the bonding between ligand and compound,
promoting dissociation and facilitate elution of the retained
compound

Immuno sorbent chromatography
Is a type of affinity chromatography, applied in immunology i.e.
antigen, anti-body purification.

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