BSA removal

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Fri Jan 14 16:04:53 EST 2000


:Nick Theodorakis wrote:
:> 
:> In article <387DF82A.BD3060FD at cajal.mbb.ki.se>, jonas at cajal.mbb.ki.se
:> (Jonas Anders) wrote:
:> > Hi all,
:> > is there a straightforward technique to remove BSA as main
:> > contaminant
:> > from the desired target protein (computed pI = 9)
:> > Any suggestions highly appreciated

Considering the very different pI of protein of interest and contaminant, 
there is no doubt that chromatofocusing will resolve them quite nicely. 
BSA does not bind to cation-exchanger at neutral pH but a protein with
pI 9 should. Thus, CM or S sepharose will also separate them. Then 
there is hydroxylapatite which has very different binding properties 
for acidic and alkaline proteins. 
 
:> I seem to recall that in the old days people used to use Blue-Sepharose
:> (Cibaron Blue?) to remove albumin from serum, since the albumin bound
:> very tightly to it but the IgG did not.
:> 
:> Nick
:> 
:> * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network *
:> The fastest and easiest way to search and participate in Usenet - Free!
:
:Cibachron/m Blue is NOT specific for albumin.  It also binds
:nucleotide-binding proteins, including heatshock proteins.
:-Han

In fact, Blue Sepharose is a _very_ non-specific sorbent and at low salt 
will bind ~ 50% of almost any given protein mixture. 

But in cases where a given protein clearly does not bind (like IgG), it 
becomes very useful. 

        - Dima





More information about the Methods mailing list