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Ion exchange chromatography

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ION EXCHANGE CHROMATOGRAPHY Ion exchange chromatography Binding of proteins depends upon pH and other ions Ion Exchange Chromatography ✓Ion exchange chromatography -- is a separation based on charge ✓Used for almost any kind of charged molecules --- large proteins, small nucleotides and amino acids ✓Ion-exchange chromatography preserves analyte molecules on the column based on ionic interactions ✓Mobile phase – buffer, pH and salt concentration--opposite charged solute ions attracted to the stationary phage by electrostatic force ✓Stationary phase– resin is used to covalently attach anions or cations onto it Principle ➢Ion Exchange Chromatography relies on charge-charge interactions between the proteins Types of IEC ➢anion exchangers ➢cation exchangers Cation exchange chromatography ---positively charged molecules are attracted to a negatively charged solid support. Commonly used cation exchange resins are S-resin, sulfate derivatives; and CM resins, carboxylate derived ions Anion exchange chromatography ---negatively charged molecules is attracted to a positively charged solid support. Commonly used anion exchange resins are Q-resin, a Quaternary amine; and DEAE resin, DiEthylAminoEthane Advantages ✓It is a non-denaturing technique. It can be used at all stages and scales of purification ✓An IEX separation can be controlled by changing pH, salt concentration and/or the ion exchange media ✓It can serve as a concentrating step. A large ...
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