ION EXCHANGE CHROMATOGRAPHY
Separates & purifies charged molecules based on their net charge.
Relies on the reversible exchange of ions between the sample molecules and charged groups immobilised on a stationary phase.
Stationary Phase: The column is packed with a matrix containing charged functional groups, either positively charged (cation exchange) or negatively charged (anion exchange).
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Sample Loading: The sample containing charged molecules is applied to the column. Positively charged molecules will bind to
negatively charged stationary phase (anion
exchange), and negatively charged
molecules will bind to positively charged
stationary phase (cation exchange).
Washing: Unbound molecules and
impurities are washed through the column using a buffer solution that doesn't disrupt the binding interaction.
To
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Elution: Elution is done by changing the ionic strength or pH of the buffer. This disrupts the electrostatic interactions between the sample molecules and the stationary phase, releasing the bound molecules.
Collecting Fractions: As different
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molecules have different affinities for the
stationary phase, they will elute at
different times, resulting in separation. Fractions containing the separated components can be collected.