Hydrophobic Interaction Chromatography

WB Lai wbl20 at cam.ac.uk
Tue Jan 30 12:55:33 EST 2001


Thank you for your response.  The problem I have is with binding the protein
on the HIC column (Phenyl).  The urea is having adverse effects on binding.
Is there a way to circumvent that, bearing in mind that the protein
self-aggregates easily?  Sorry I haven't been clear in my message.

H.V.Taylor wrote in message <7eN2OnDKOI+OfGeMjpIP84QZNn4a at 4ax.com>...
>
>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
>error...
>
>Harold
>







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