11.24 : アフィニティークロマトグラフィー

Affinity chromatography is a separation technique to purify biomolecules from complex mixtures.

The separation is driven by a biospecific binding interaction between an analyte and its affinity ligand, such as antigens and antibodies, enzymes and substrates, or receptors and hormones.

The affinity ligand is in the stationary phase, being immobilized on a support such as agarose or porous glass beads, which are packed into a column.

The mixture containing the analyte is in the mobile phase, which must provide the right conditions for the binding interaction.

As the mixture passes through the column, the analyte selectively binds to the affinity ligand while the other molecules pass through the column.

Afterwards, the bound analyte is eluted by changing the mobile phase conditions, such as ionic strength or pH, to weaken the binding with the ligand and therefore obtaining the purified analyte.