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Sample
Calculation:
Purification Factor


The purification factor (also called enrichment) is a comparison of the specific activity at some point in the purification process to the specific activity at the beginning of the purification. The formula for purification factor is as follows:

Specific activity of separation step
------------------------------------------
Specific activity of crude extract


Since the specific activity should increase with each separation step, the purification factor should always be greater than 1. The purification factor has no units since the units of specific activity are in both the numerator and denominator and will cancel out.

For example, the protein concentration for the crude extract may be 1000 mg/mL and the enzyme activity might be 30,000 units/mL, leading to a specific activity of 30 units/mg.

After the second step, the protein concentration may be 100 mg/mL and the enzyme activity could be 25,000 units/mL. So the specific activity would have improved from 30 units/mg to 250 units/mg. Although you may have lost some, you removed a lot of contaminating proteins so the specific activity improves from 30 units/mg to 250 units/mg.

To express this as a purification factor:

250 units/mg = 8.33
----------------
30 units/mg

So, the purity has improved by a factor of 8.33. Or, to say it a different way, the protein is 8.33 times more pure than it was when you started.

 

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Contact Us:

Lisa Seidman
lseidman@matcmadison.edu
(608) 246-6204

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