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Re: [ccp4bb] FW: endotoxin removal

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Subject: Re: FW: endotoxin removal
From: lieh low liehyoon {- at -} GMAIL {- dot -} COM
Date: 2012-03-07

Hi Jerry,
We have used 0.05% Triton X-100 in our wash buffer to wash the protein
on column (Ni-IMAC for His-tagged proteins and MabSelect for
antibodies). 20 column-volumes wash followed by 20 column-volume wash
without detergent is typically enough to reduce the endotoxin
significantly.
I would also suggest checking your FPLC system, and decontaminate with
2N NaOH if neccessary.
Hope that helps
Regards,
ray

On Wed, Mar 7, 2012 at 9:50 AM, Jerry McCully
wrote:
>
>
> Dear All;
>
>         We purified a His tag protein by Ni-NTA and gel-filtration from
> E.coli.
>
>          We tried two endotoxin removal resins from Pierce. However, it is
> hard to remove the endotoxins in the purified protein because the protein
> bound to the resin as well.
>
>          This protein contains quite a few aromatic residues and has a pI
> around 6.
>
>          Any ideas to remove the endotoxins will be highly appreciated.
>
>         best regards,
>
> Jerry McCully
>
>




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