To compare the effectiveness of various types of column chromatography we use a series of parameters. Which one of the following would be the best to compare between purifications schemes?

Question

BlackTom AI · Accepted Answer

In evaluating which parameter best compares purification schemes, consider what each metric actually measures and how it reflects both purity and activity.
Option 1: Total protein. This measures how much protein is present, but it does not indicate how much of that protein is the desired, active enzyme. A purification step could remove contaminating proteins but leave total protein high if the target is lost or degraded, making total protein a poor standalone measure of purification success.
Option 2: None of the above. This would be selected only if none of the listed metrics were useful, which is not the case here because there is a widely accepted parameter that effectively compares purification outcomes.
Option 3: Specific activity. This is defined as enzyme activity per unit of protein (e.g., units per mg). It directly reflects both the activity of the target enzyme and the purity of the preparation, allowing meaningful comparison of purification efficiency across schemes. A higher specific activity indicates a purer, more active preparation, which is precisely what one wants to optimize when purifying.
Option 4: Yield. Yield measures the amount of product recovered relative to the starting material but does not account for purity or activity. A purification step could have high yield but very low purity or vice versa. Therefore, yield alone can be misleading when comparing purification schemes.
Thus, among the options, specific activity best serves as a basis for comparing purification strategies, because it balances activity with amount of protein and directly reflects purity improvements across steps.

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To compare the effectiveness of various types of column chromatography we use a series of parameters. Which one of the following would be the best to compare between purifications schemes?

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