CHROMATOGRA

PHY
ION EXCHANGE
CHROMATOGRAPHY
 Ion exchange chromatography (IEC) is a widely used technique for protein purification

because of its
 versatility,

 high resolving power,

 high capacity

 simplicity in operation.

 The basic principle in IEC involves reversible competitive binding of different ions of one

kind to immobilized ion exchange groups of opposite charge bound to the chromatographic
matrix called the ion exchanger.
 When a sample mixture containing components of positive charge is applied to a column

containing cationic (negatively charged) matrix, solute components with more positive
charges (M2+) are adsorbed (exchanged) strongly compared to those with less positive
charges (M+).

 Components with no net charge (M0) or a net negative charge (M-) pass through the column

unretained.
 The interaction between the proteins and the ion exchanger depends on several factors such

as
 the net charge and charge anisotropy - charge distribution on the protein surface,

 ionic strength and pH of the solvent,

 nature of ions and other additives.

 The higher the charge on the protein, the more strongly it will bind to a given ion exchanger

of opposite charge.

 The pH of the medium or solvent is the most important parameter, which determines protein

binding, as it determines the effective charge on both the protein and the ion exchanger.