Affinity chromatography is a method developed in 1930 by Swedish biochemist Arne Tiselius to study enzymes and proteins. It relies on the affinity between biochemical compounds and specific properties. Affinity chromatography separates biochemical mixtures using highly specific interactions between antigens and antibodies, enzymes and substrates, or receptors and hormones. It is used to purify and concentrate substances from mixtures through buffering solutions and can reduce substance amounts in mixtures.

1. Affinity
Chromatography

2. ∗1930, First developed by A.Wilhelm Tiselius,
Swedish biochemist, won the Nobel Prizein
1948
∗Used to study enzymes & other proteins
∗Relies on the affinity of various biochemical
compounds with specific properities.
Affinity History

3. ∗A method of separating biochemical
mixtures based on a highly specific
interaction between antigen & anti-
body, enzyme & substrate, or receptor
& hormone.
Affinity
Chromatography

5. Antigen Antibody
Enzyme Substrate
DNA Histon
Examples:

6. ∗Can be used to:
∗Purify & concentrate a substance from a
mixture into buffering solution.
∗Reduce the amount of a substance in
mixture
∗Purification of IgG fragments.
Uses:

7. ∗ The sample is injected into the equilibrated
affinity chromatography column.
∗ Only substance with affinity for the ligand are
retained on the column.
∗ The substance with no affinity to the ligand will
elute off.
Mechanism of Affinity
Chromatography:

10. ∗ Used in genetic engineering , ex: nucleic acid
purification.
∗ Production of vaccines , ex: anti-body purification
from blood serum.
∗ Basic metabolic research , ex: protein or enzyme
purification from cell.
Application:

13. ∗Extremely high specificity.
∗High degrees of purity can be
obtained.
∗The process is very reproducible
∗The binding sites of biological
molecules can be simply
investigated.
Advantages of Affinity
Chromatography:

14. ∗Expensive ligands.
∗Leakage of ligand.
∗Degradation of the solid support.
∗Limited lifetime.
∗Non-specific adsorption.
∗Relative low productivity.
Disadvantages:

15. ∗Hanady Khaled
∗Hend Ahmed
∗Hend Batea
∗Hend Gamal
∗Hend Hassan