Your research project requires that you purify protein A away from proteins B an
ID: 1058925 • Letter: Y
Question
Your research project requires that you purify protein A away from proteins B and C using a combination of gel filtration and ion exchange chromatography. Describe a purification scheme using just these two columns separately that would accomplish your goal with the least amount of protein dilution. Induce in your answer the expected order of elution of the proteins from the two columns, given the physical properties of the proteins in the following table. Also, justify your answer giving the scientific reasons behind your scheme.Explanation / Answer
Separation of proteins A, B and C
Protein A and C have relatively same molar mass but bery different isoelectric point. So the two proteins are separated by adjusting the pH by gel electrophoresis. This technique is dependent upon the overall charge on the protein as a function of pH of the gel.
Protein B has a very different molar mass and can be separated from protein A, by ion-exchange chromatography. Starting from a low pH we move to high pH and depending upon the net charge on the protein, they exchange with the ion-exchange resin and get adhered to it which is later eluted out at higher pH value.
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