affinity

Kiley R. Prilliman kprillim at rex.re.uokhsc.edu
Wed Apr 9 23:00:33 EST 1997


> hey ,
> 
> I was wondering why you did not use a proteine A/G crosslinked
> monoclonal instead of CNBR sepharose crosslinked. I don't think the
> small difference in the pH explain your problem! 
> 
> didier

Well, back a year ago when we were starting to develop an affinity
column for our purification protocol, we experienced some difficulties
with protein A or G matrices (I would have to go back through others'
notebooks to identify the problems specifically, and I am not at the lab
right now), and much of the literature regarding purification of the
protein we are looking at refers to CNBr matrices.

Incidentally, as insane as this might sound, I re-ran the "old" matrix
(coupled last year) with protein-containing supernatant set to pH 7.0
today, and it worked.

(Maybe it was the pH, or maybe it was the sacrificial voodoo ritual I
performed to the affinity chromatography gods, who knows)...

Kiley R. Prilliman
Department of Microbiology & Immunology
University of Oklahoma Health Sciences Center

phone: 405-271-1203
  fax: 405-271-3117



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