Ion Exchange Column Chromatography

For interaction to occur the protein of interest must have a charge opposite to that of the functional group of the sorbent particle.

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. Key steps in the ion exchange chromatography procedure are listed below. An impure protein sample is loaded into the ion exchange chromatography column at a particular ph. It can be used for almost any kind of charged molecule including large proteins small nucleotides and amino acids.

It is usually used for protein purification but may be used for purification of oligonucleotides peptides or other charged molecules. 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. Ion exchange chromatography workflow. Ion exchange chromatography is an interesting type of column chromatography.

Mobil phases consist an aqueous buffer system into which the mixture to be resolved. 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. 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. Ion chromatography or ion exchange chromatography separates ions and polar molecules based on their affinity to the ion exchanger.

This separation is done based on the differences in the adsorption coefficient or partition coefficient of the sample with the stationary phase. Ion exchange chromatography separates molecules based on differences between the overall charge of the proteins.