On the DEAE, try lower ionic strength or slightly higher pH,
try DEAE-Sepharose instead of cellulose, to get it to stick
Try a cation exchange column like carboxymethyl- or phospho-
cellulose; at slightly acidic pH if necessary.
eab
Narayanan Ramasubbu wrote:
> Hi all:
> I am trying to purify a protein that was expressed in a baculovirus
> system. The protein elutes in the flow through in a DE52 column. Further
> gel filtration gives an enriched protein but we see that some medium
> components tag along (weight and activity data do not match). Another
> mutant in the series also gives 30 mg of colorless fluffy material but
> again we never got this much protein for other mutants in the same
> series. We suspect that some "medium" is tagging. We tried other
> chromatography such as hydroxyapatite, activated charcoal, anion
> exchange etc but not "successful" in getting rid of the "impurity".
>
> Has anyone come across a protein that eluted in the flow through and if
> so, what is their experience? Did they notice such behavior?
>
> Your advice will be greatly appreciated.
>
> Thanks
> Subbu
>
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