How to remove BSA from protein sample? - (Aug/01/2006 )

Hi collegues!

i m currently performing protein purification (preparative scale) and unfortunately, my target protein represent a very very little ratio from all proteins (why it is always like that ?!) and bsa is the main protein from my samples.
so my first goal is very simple: to find a way to remove bsa in a very efficient way so that I could more easily purify my target protein with hplc. I would not go on immunoprecipitation or things with immunoaffinity against bsa (too expensive regarding the quantity of bsa) but if anybody know a procedure before or during hplc to specifically discard bsa, or even litterature references, it is very welcome.

thank all

Dj

Thanks Chakchel for your suggestion.
Blue sepharose is something that I will try but I m quite sure that it will also bind my target, but I agree that it worth to be try. If u got other tips, you are welcome

dj

QUOTE (Djin @ Aug 1 2006, 11:14 AM)

Hi collegues!

i m currently performing protein purification (preparative scale) and unfortunately, my target protein represent a very very little ratio from all proteins (why it is always like that ?!) and bsa is the main protein from my samples.
so my first goal is very simple: to find a way to remove bsa in a very efficient way so that I could more easily purify my target protein with hplc. I would not go on immunoprecipitation or things with immunoaffinity against bsa (too expensive regarding the quantity of bsa) but if anybody know a procedure before or during hplc to specifically discard bsa, or even litterature references, it is very welcome.

thank all

Dj

Hi!

Well, I never did that, but a friend of mine used Blue-Sepharose to remove BSA, that binds very selective and tightly. Other proteins didn't...
Maybe you should check if you get some information about this method (or maybe I will ask him the next time I see him ).

Chakchel

http://www4.amershambiosciences.com/aptrix...nline_issue_9_9

I dont know if you can use it for practical reasons for prep scale application though. Have you tried ion exchange column?

Hi, it's me again!

I talked to my friend yesterday and he said that it would be very important to test different pHs for the removal. He remembered that for BSA he used a 20mM PBS-solution as binding buffer (pH 7,2).

Hope this will help you!

Chakchel

thanks for your interest
what sort of procedure was following the 20mM PBS pH7.2 for discarding bsa, iec or blue seph?
i currently triing iec and tuning pH looks very interesting for removing bsa.

dj

thanks for your interest
what sort of procedure was following the 20mM PBS pH7.2 for discarding bsa, iec or blue seph?
i currently triing iec and tuning pH looks very interesting for removing bsa.

dj

Currently there is several commercially available kits for this purpose, but as already know you are going still lose your low abundant protein as you already know. One kit I had good success with fro a similar purpose I have bought from Norgen Biotek and basiclly is for removal of high ubandant serum proteins. The nice thing about that thier technology is pI dependent (i.e. if your protein has an acidic pI you will enrich for it and they have altrante procedure where if you have a basic protein you can enrich for it as well!

Good luck!