Biochemical Techniques and Instrumentation

2.4.4. Immobilization methods

Ligands are coupled to the support matrix by a series of reactions. The reactive side chains of the support matrix are chemically modifed before the attachment of the ligand by different activation methods such as cyanogen bromide, triazine, periodate oxidation, epoxide, acyl azide, diazo, succinate esters, carbodiimide and carbonylation.

All the reactive side chains in the support matrix are not coupled to the ligand. There are uncoupled side chains that can bind with specific and non-specific components. Blocking agents are used to block the uncoupled side chains. eg. Human IgG is used as a blocking agent to isolate human IgE.