Hydrophobic Interaction Chromatography
harold.taylor.remove-this at delete-this.uni-tuebingen.de
Tue Jan 30 11:29:42 EST 2001
"WB Lai" <wbl20 at cam.ac.uk> wrote:
>I am trying a difficult purification of a denatured protein (suspended in 8
>M urea) on a HIC column. Has anyone had any success with HIC purification
>in the presence of urea?
>My main problem is that the protein aggregates severely in chaotrope
>concentration < 5M urea.
>Any help is greatly appreciated.
What type of HIC are you using? If it's C4-, C8- or phenyl- you could
try adding organic solvents to elute (your protein is denatured
anyway). I'd start with glycerol or ethylene glycol as they will more
likely be compatible with >5M urea than other solvents. However, the
self-aggregation of your protein may be suppressed by glycerol or
ethylene glycol as well. As usual, it's a question of trial and
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