Eluting mAb rat IgG from ascites

Robert Burns burns at sasa.gov.uk
Fri Aug 26 04:10:52 EST 1994


"P.M.Grandison" <damian.white at genesis.co.nz> wrote :

>Greetings.
>Im having trouble eluting monoclonal rat IgG from ascites fluid using
>protein G HiTrap afinity columns ( 1mL volume ) - there seems to be a 
high
>back pressure causing the column to block up.
>Has anyone out there experienced this?  If so, any solutions . . ?
>
>P. M. Grandison
><prudence.grandison at genesis.co.nz>
>Genesis Research & Development Corporation
>P.O.Box 50
>Auckland
>New Zealand
>
>
>

Hi,
I would imagine that your column is blocked with debris from your
ascites. We always put everything through a 0.2u filter prior to
putting on to the column.

You could try attaching a syringe via a thin piece of tubing to the
outlet end of the column and flushing it through backwards to get
rid of the debris. Hope that this helps


Robert Burns
Scottish Agricultural Science Agency
Edinburgh
Scotland

burns at sasa.gov.uk




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