Ion Excnage Chromatography

[mariacervantes888333]

 

In ion exchange chromatography, how do you find the order of elution of proteins based on pI?

 

I have a graph of KCl conc vs. fraction number and I have to find the conc of KCl required to elute some given proteins.

 

And I'm given pI values of 4.6-4.9 for albumin and 6.8-7.8 for hemoglobin, and 10 -11 for cytochrome c, so what do I do with these numbers in relations to my KCl conc std curve and my ionic conductivity of fractions?

[DeltaScience]

Hello!, well I dont know much about ion exchange cromatography but i do know that the isoelectric point of a protein determines it charge, and in this case it dependes basically on the pH of your elution buffer, for instance, for albumin (4,6-4,9) if you decrease the pH to a point lower than 4,6; then the protein will elute, and so on for the other proteins, also since in this case you're using KCL, you can try by using different pH or concentrations, the more the difference between the pi of a protein and the buffer you're using the more concentration of KCL you'll need to elute it.

 

here have a look at this i found on the web, I hope its usefull for you: http://www.piercenet.com/files/TR0062-Ion-exchange-chrom.pdf

 

I know it might not be the answer you're looking but hopefully will give you more usefull information.

 

Regards