Ion Exchange Column Chromatography

As you know the chromatography is a process of the separation of molecules from a mixture.

Ion exchange column chromatography.

Ion exchange chromatography separates molecules based on differences between the overall charge of the proteins. Pierce ion exchange spin column purification example. Working principle of ion exchange chromatography. Ion exchange chromatography is widely used for separating diverse compounds including carbohydrate and alcohol isomers as well as sugar phosphates and nucleotides.

However there are also thin layer chromatographic methods that work basically based on the principle of ion exchange. Anion exchangers are used at ph values above the isoelectric point of the protein where the net charge on the protein is negative. Ion exchange chromatography is most often performed in the form of column chromatography. Key steps in the ion exchange chromatography procedure are listed below.

Column cleaning for ion exchange chromatography and chromatofocusing practical considerations for iex separation sepharose big beads. Tr0062 0 ion exchange chromatography. Therefore use a purification buffer at ph 5 2 for the anion q columns or a ph 5 2 for the cation s columns. Ion exchange chromatography iec a method of separating molecules such as proteins on the basis of their net charge ion exchange columns may have either positive or negative groups giving anion or cation exchangers respectively.

For interaction to occur the protein of interest must have a charge opposite to that of the functional group of the sorbent particle. Example protein x has a pi of 5 2. An impure protein sample is loaded into the ion exchange chromatography column at a particular ph. Ion exchange chromatography workflow.

Purification from crude viscous samples at large scale. It is usually used for protein purification but may be used for purification of oligonucleotides peptides or other charged molecules. 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 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.