Despite the potential of any other techniques Ihave been forced to use a combina

Question

Despite the potential of any other techniques Ihave been forced to use a combination of size exclusion chromatograghy and ion exchange chromatograpthy Which of the following statements must have been true? Pick ALL that apply A. B. I would use an acetonitrile mobile phase with a pH of 7.4 I would use a water based mobile phase of pH 7 2 D. I would use an acetonitrile phase of 7.2 E. I would salt it out. F I would obtain my soluble protein fraction in native conformation by dialysis to remove salts or to modify the pH G would obtain my soluble protein fraction by eluting orn he column in stages slowly changing the pH

Explanation / Answer

When size exclusion is being used, it means the separation takes place based on the charge where smaller ones get stuck. In case of ion exchange the separation is based on the ionic and electric charge. Hence in this case the answer will be b, e, g.

It is because acetonitrile can be used at pH 7.4 and it helps in separation of the proteins efficiently. Salting out again is a mechanism where through aggregation there is separation. Chnagng the pH slowly helps in dissolving the protein of interest and hence pH needs to be adjusted accordingly

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