Affinity chromatography

Affinity chromatography

1. what is it?

   • Enables the purification of a biomolecule with respect to biological function or individual chemical structure.

2. used for?

   • Monoclonal and polyclonal antibodies
   • Fusion proteins
   • Enzymes

3. concept?

   • A solvent is used for carrying sample mixture through the matrix.
   • Separation occurs because each compound in a mixture interacts differently with the matrix.

4. steps?

   • Equilibration
     • Sample application
       • Binding and washing
         • Desorption and elution

5. columns used?

   • Ni2+
   • Specific antibodies
   • glutathione
   • Protein A or G

6. concept?

   • A solvent is used for carrying sample mixture through the matrix.
   • Separation occurs because each compound in a mixture interacts differently with the matrix.

7. IMAC

   • Immobilized Metal ion Affinity Chromatography
     • takes advantage of the ability to histidine residues to interaction with various transition metal ions