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The following is the chromatogram of an anion exchange column. The absorbance, v

ID: 489648 • Letter: T

Question

The following is the chromatogram of an anion exchange column. The absorbance, volume of elution, and the salt gradient concentration are shown in the different axes. Two proteins, A and B were purified using the column. Answer the questions based on the chromatogram.

Charge of A:

Charge of B:

Which protein has more net charge to bind to the column tightly?

What salt concentration did A elute at?

What salt concentration did B elute at?

Which of the two proteins are bigger?

0.6 M 0.4 M 8 0.2M 15 25 35 55 65 75 85 Volume (mL) uonenueou00 IOeN MM B A 5 08ZV

Explanation / Answer

CHARGE OF B IS + AS IT WASHED OUT AND CHARGE OF B IS -VE PROTEIN A HAS ANETCHARGE TO BIND TO THE COULUMN SALT CONCENTRATION FOR A IS 20 AND SALT CONCENTRATION OF B IS 50 AS PER SALT CONCENTRATION PROTEIN B IS BIGGER