Ion Exchange Column Chromatography

An impure protein sample is loaded into the ion exchange chromatography column at a particular ph.

Ion exchange column chromatography.

Ion exchange chromatography is an interesting type of column chromatography. After loading an impure protein sample onto an ion exchange chromatography column the column is washed to remove undesired proteins and other impurities and then the protein s of interest is eluted using either a salt gradient or a change in ph. For interaction to occur the protein of interest must have a charge opposite to that of the functional group of the sorbent particle. It can be used for almost any kind of charged molecule including large proteins small nucleotides and amino acids.

Key steps in the ion exchange chromatography procedure are listed below. Ion exchange chromatography separates molecules based on differences between the overall charge of the proteins. This separation is done based on the differences in the adsorption coefficient or partition coefficient of the sample with the stationary phase. As you know the chromatography is a process of the separation of molecules from a mixture.

Ion exchange chromatography workflow. Mobil phases consist an aqueous buffer system into which the mixture to be resolved. It is usually used for protein purification but may be used for purification of oligonucleotides peptides or other charged molecules. Ion chromatography or ion exchange chromatography separates ions and polar molecules based on their affinity to the ion exchanger.

It works on almost any kind of charged molecule including large proteins small nucleotides and amino acids however ion chromatography must be done in conditions that are one unit away from the isoelectric point of a protein. Ion exchange chromatography definition or ion chromatography is a process that allows the separation of ions and polar molecules based on their affinity to the ion exchanger.