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Biochemistry

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Biochemical Protocol: DEAE Ion Exchange Chromatography

BIOCHM-81FILH

You and your lab partner are assigned to purify an enzyme from a yeast extract. Everything is proceeding well until your lab partner forgets to dialyze your sample after batch DEAE (diethyl-aminoethyl)-ion exchange chromatography in which NaCl was used to elute the protein.

Worse, they did not tell you this before you carried out the column DEAE-ion exchange chromatography protocol.

Which of the following statements describes the MOST likely result of this change in the protocol?

A

Failure to dialyze the extract would not affect binding or elution of the protein in the column DEAE-ion chromatography step, but it also would not lead to an increase in the purification of the protein.

B

Failure to dialyze the extract likely resulted in the protein of interest not binding to the resin in the column DEAE-ion chromatography step; rather, it passed through in the initial application and wash.

C

Failure to dialyze the extract would likely result in denaturation of the enzyme prior to the column DEAE-ion exchange step.

D

Dialysis of the extract is not an essential step between the two ion-exchange procedures.

E

Failure to dialyze the extract likely resulted in the protein of interest binding to the resin in the column DEAE-ion chromatography step, but not subsequently eluting.