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Suppose that you are given a mixture of proteins with their properties provided in the following chart.

Isoelectric point PI Molecular weight
Protein A 4.1 80
Protein B 9.0 81
Protein C 8.8 37
Protein D 3.9 172
Choose one combination of techniques that can be used to isolate Protein B from Proteins A, C, and D.
a. gel filtration chromatography and ultracentrifugation
b. dialysis and ultracentrifugation
c. dialysis and ion exchange chromatography
d. ion exchange chromatography and gel filtration chromatography

1 Answer

8 votes

Answer:

The correct option is d. ion exchange chromatography and gel filtration chromatography

Step-by-step explanation:

The mixture contains proteins with different isoelectric points and molecular weights. Ion exchange chromatography separates the proteins with different electric charges, which depend on the isoelectric point. At certain pH of the aqueous medium, the proteins will have a certain electric charge and they will be retained differently in the stationary phase. Proteins A and D have similar isoelectric points (around 4.0) and maybe they cannot be efficiently separated by this method; the same occurs with protein B and C (with an isoelectric point around 9.0). To separate proteins A from D and protein B from C, it could be used gel filtration chromatography. This separation method is based on differences in molecular weights. So, at the end of the process, we will obtain the proteins A, B, C, D separated one from each other.

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