Protein purification

[SamP]

So I have a "hypothetical" worksheet for biochemistry. I have to separate the following 8 proteins as much as possible:

 

Bovine Serum Albumin

Myoglobin

Hemoglobin

Cytochrome C

Alpha-lactalbumin

Ribonuclease

Ferritin

Catalase

 

 

I started with Size-exclusion and got these for the light proteins:

Cytochrome C

M`yoglobin

Alpha-lactalbumin

Ribonuclease

 

And heavy proteins:

Hemoglobin

BSA

Ferritin

Catalase

I figured out ways to separate the light proteins further and essentially have them completely separated. What are some techniques I can use to separate the heavy proteins?

Edited March 26, 2015 by SamP

[CharonY]

There are so many around, I guess it depends what has been covered in your course so far. A few parameters to think about: pI, charge, hydrophobicity, mw (already partially covered). There are a few techniques which allow pretty decent separation of all of them in one workflow.

[Vexen]

Cation Exchange

 

Anion Exchange

 

Affinity Separation

 

If you use immunoaffinity, you could separate in one step.

 

Reverse phase chromatography could also be useful.

There are various affinitychromatographic columns that have affinity for specific groups of proteins.

 

If you know the molecular mass, you could just use SDS-Page.