Ion Exchange Column Chromatography

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 column chromatography.

Ion exchange chromatography separates molecules based on differences between the overall charge of the proteins. Key steps in the ion exchange chromatography procedure are listed below. For interaction to occur the protein of interest must have a charge opposite to that of the functional group of the sorbent particle. 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 exchange chromatography is an interesting type of column chromatography. This separation is done based on the differences in the adsorption coefficient or partition coefficient of the sample with the stationary phase. An impure protein sample is loaded into the ion exchange chromatography column at a particular ph.

Ion exchange chromatography workflow. As you know the chromatography is a process of the separation of molecules from a mixture. Because of donnan exclusion ionic material is excluded from the ion exchange resin and passes quickly through the column. Ion chromatography or ion exchange chromatography separates ions and polar molecules based on their affinity to the ion exchanger.

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. It can be used for almost any kind of charged molecule including large proteins small nucleotides and amino acids.