3) DEAE Sepharose is a weak anion exchange resin containing the positively charg

Question

3) DEAE Sepharose is a weak anion exchange resin containing the positively charged functional group6ther -CHyCH2NH'(C2H5)2-1f DEAE Sepharose resin is used to separate three different proteins (#1 is positively charged #2 is eaky negatively charged and#3 is strongly negatively charged), which protein(s) would be in the flow-through? Which protein(s) would be bound in the resin? If two or more points) proteins are bound in the resin, what would be the order of elution in an increasing NaCI gradient? (5

Explanation / Answer

DEAE Sepharose resin contains positively charged functional groups that interact favourably with the negative charges. So the positively charged proteins present in the flow through, while the both the weakly and strongly negatively charged protein bound to the resin.

Addition of NaCl will replace the negatively charged protein by the Cl- group ( Cl- interact with positively charged functional groups of the resin. So the weakly negatively charged proteins elute first followed by the strongly negatively charged protein as NaCl concentration increases.

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