Ion Exchange Column Chromatography

Ion exclusion chromatography is defined 91 as a technique used to separate weak acids amino acids sugars alcohols and other substances on an ion exchange column.

Ion exchange column chromatography.

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. Mobil phases consist an aqueous buffer system into which the mixture to be resolved. Key steps in the ion exchange chromatography procedure are listed below. 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 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 separates molecules based on differences between the overall charge of the proteins. Ion exchange chromatography is an interesting type of column chromatography. An impure protein sample is loaded into the ion exchange chromatography column at a particular ph.

This separation is done based on the differences in the adsorption coefficient or partition coefficient of the sample with the stationary phase. For interaction to occur the protein of interest must have a charge opposite to that of the functional group of the sorbent particle. Because of donnan exclusion ionic material is excluded from the ion exchange resin and passes quickly through the column. As you know the chromatography is a process of the separation of molecules from a mixture.

Ion exchange chromatography which is designed specifically for the separation of differently charged or ionizable compounds comprises from mobile and stationary phases similar to other forms of column based liquid chromatography techniques 9 11. Ion exchange chromatography workflow.