The Physical Properties of Two Isoforms of a Protein, Both

The physical properties of two isoforms of a protein, both of which bind the same ligand, are shown in the table: Which protocol would yield the best separation of the two isoforms?

A) Run the mixture through a cation-exchange column starting at pH 10, then gradually increase the pH of the buffer.
B) Pass the mixture through a size-exclusion column with a pore size cutoff of 40 kDa.
C) Load the mixture onto an anion-exchange column at physiological pH, then gradually increase the NaCl concentration of the buffer.
D) Load the mixture onto an affinity column with the ligand covalently bound to the beads, followed by cleaving the covalent bond to elute.

Correct Answer:

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