> 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@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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