"WB Lai" <wbl20 at cam.ac.uk> wrote:
: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.
Naturally, since denaturing action of urea is disruption of hydrophobic
interactions. If you have to keep your protein in 5 M urea and under
this conditions it does not bind to phenyl sepharose, then it means
phenyl sepharose is not suitable for you. I don't see a way around
You can try a different sorbent. Octyl or even Dodecyl Sepharose
will bind a lot better than Phenyl in HIC. Maybe the strenght of
binding will be just enough for your purpose.