IPA precipitation of virus

Morten Kring mortenkring at hotmail.com
Wed Apr 17 03:44:51 EST 2002


Ok, but...... what about desalting. I mean, after the precipitation
I need to make a dialysis on the supernatant or run it on a G-25 column and
thereby risking
a big loss in specific activity ?
I am well aware that protein purification isn't achievable without a loss in
every step. Nothing is free.
But I am afraid that the desalting step will be too much...

"D.K." <dk at no.email.thankstospam.net> wrote in message
news:a9ieh4$g6b$1 at news.doit.wisc.edu...
> "Morten Kring" <mortenkring at hotmail.com> wrote:
> >That is of course an alternative. But what about pH of the solution ? and
> >roughly how low is low % ammonium sulphate?
>
> pH is neutral (and preferably well-buffered as AS acidifies a bit)
> and I would start try 5, 10, 15, 20% saturation as proteins do
> not precipitate significantly at this range.
>
>
> >"D.K." <dk at no.email.thankstospam.net> wrote in message
> >news:a9h4k0$qb7$1 at news.doit.wisc.edu...
> >> "Morten Kring" <mortenkring at hotmail.com> wrote:
> >> >I'm looking for help and hints on how to remove virus from a
> >> >ionexchanger eluate containing blood plasma proteins. In particular
i'm
> >> >looking for methods
> >> >to remove MVM (minute virus og mice) which is a part of the
Parviviridiae
> >> >familiy and is especially
> >> >difficult to handle.
> >> >I was thinking of doing a fractionated precipitation with IPA
> >> >(isopropyl-alcohol or isopropanol) at around 25 % vol./vol. IPA. Does
> >anyone
> >> >know if this could remove or inactivate the virus at this relatively
low
> >> >concentration of organic solvent?
> >>
> >> What about low % of ammonium sulphate? Viruses should salt out
> >> readily.
> >>
> >> DK
> >
> >





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