The following data were obtained by the authors while purifying the protein desc

Question

The following data were obtained by the authors while purifying the protein described in question 1 (OfHex). Three different columns were used in sequence. All fractions were analyzed for protein content (by A^280 measurements; closed symbols and black line) and for OfHex catalytic activity (A^405; open symbols and grey line). Conductivity measurements were also taken to show that buffer gradients had been created (Cond (mS-cm^-1). Place the chromatograms in proper order, and explain your reasoning.

Explanation / Answer

The proper order will be A > B > C.

On the first chormatogram, researchers are getting a high activity (fractions 4-7) but also the have a large amount of proteins. This means that the protein isn't properly purified. On the second chromatography, fractions 4-7 where pased thru a new ionic exchange column. This allowed to separate to peaks: the first one for other proteins and the secon one will carrye the higher amount of the proteins of interest (fractions 18-26).

Last, the fractions 18-26 where pased thru a third collumn, for getting the maximun rate of purification. In C chromatogram you can see that the two peaks (for enzyme activity and protein concentration) are whell separeted. That means that the enzyme is correctly purified.

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