Which purification protocol is the best?

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Nov 10, 2017
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Given purification of the same enzyme using 4 diffrerent protocols, based on the concentration, volume and total activity of enzyme from each protocol, which is the best? What do i use as an indicator as to which protocol is the best? Specific enzyme activity or yield?

Protocol A: 100uL, 20mg/mL, Total activity: 100units/s

Protocol B: 100uL, 50mg/mL, Total activity: 10units/s

Protocol C: 500uL, 20mg/mL, Total activity: 100units/s

Protocol D: 100uL, 10mg/mL, Total activity: 1000units/s

5 Comments

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relaxin 11 months ago

In general, the higher specific activity the better. With high yield and low specific activity, you may lose more when you have to purify the sample with additional steps. It looks like protocol D gives highest specific activity.

What do you mean by 1000 units/s? Specific activity is expressed as units/ug protein.

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exia1995 11 months ago

Im nt very sure but the question was just set like this and asked us which protocol is the best based on the results of the enzyme for each of them. Looks to me like production of product? I chose D too on the basis of activity

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mdfenko 11 months ago

simply put, total activity is the amount or yield of enzymatic activity, specific activity is the purity of the enzyme.

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exia1995 11 months ago

So would Protocl D be the best answer?

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mdfenko 11 months ago

protocol d gives the highest specific activity (best purity). that's what i would go for.