Problems with monoclonal antibodies

bourin bourin at attglobal.net
Fri Sep 1 04:43:12 EST 2000


> You may try alternative IgG purification protocols. One works relatively fine, and it
> uses very soft elution conditions. It is thiophilic gel chromatography (Sigma sell this
> Gel). You could found usefull protocols in

Lihme et al. (1991) Anal. Biochem. 192, 64-69
Joubert-Caron et al. (1994) Int. J. Biochem. 26, 813-823

Good luke,

Dr Bourin



>  ----------
> From: A.Smith at centenary.usyd.edu.AU
> To: cyto-inbox
> Subject: Problems with monoclonal antibodies
> Date: Wednesday, August 30, 2000 20:58
>
> Hi all,
>      We have been having some problems with some of our
> monoclonals and would appreciate any advice or comments...
>
> Bascially we have two problems which may be related:  after
> purification on Protein A or G and elution (glycine pH2.5 then tris
> pH8 neutralisation and dialysis against PBS), the antibody either
> precipitates spontaneously at 4 degrees, or the biotinylated
> conjugates rapidly lose activity (which may also be due to
> precipitation - we haven't checked yet). We suspect low levels of
> acid denaturation may cause ongoing spontaneous formation of
> aggregates. Does anyone have any experience of this and how to get
> around it (most mAbs are fine after purification this way and only
> some mAbs seem to be affected)?
>
> Thanks,
> Adrian
>
>  --
> ******************************************************
> Adrian Smith (PhD Student)      T CELL BIOLOGY GROUP
> Centenary Institute of Cancer Medicine & Cell Biology
> Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.
> ******************************************************
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