calculating yield
Posted: Fri Sep 11, 2015 11:46 am
hi all,
i'm running a capture anion exchange step on an akta on filtered harvest producing an enzyme.
i just want to double check my logic in calculating the yield is correct:
I feed in the column 7.5ml of harvest containing 0.06mg/ml of product = 0.45mg (product NOT total protein)
I collect 2ml fractions throughout the whole chromatogram.
my eluate peak comes out as 6 fractions (12ml) and the concentration in each fraction varies between 0.01 to 0.04 mg/ml. I averaged out the concentrations to 0.025mg/ml, therefore 12 *0.025 = 0.3mg
my yield is therefore 0.3/0.45 x 100 = 67% and i lose 33%.
question 1) is this the yield or the protein recovery?
question 2) shouldn't i see a more concentrated product in the eluate?
question 3) does this make sense to you?
thank you for your help
i'm running a capture anion exchange step on an akta on filtered harvest producing an enzyme.
i just want to double check my logic in calculating the yield is correct:
I feed in the column 7.5ml of harvest containing 0.06mg/ml of product = 0.45mg (product NOT total protein)
I collect 2ml fractions throughout the whole chromatogram.
my eluate peak comes out as 6 fractions (12ml) and the concentration in each fraction varies between 0.01 to 0.04 mg/ml. I averaged out the concentrations to 0.025mg/ml, therefore 12 *0.025 = 0.3mg
my yield is therefore 0.3/0.45 x 100 = 67% and i lose 33%.
question 1) is this the yield or the protein recovery?
question 2) shouldn't i see a more concentrated product in the eluate?
question 3) does this make sense to you?
thank you for your help