Problems with monoclonal antibodies

From: Adrian Smith (A.Smith@centenary.usyd.edu.AU)
Date: Tue Aug 29 2000 - 20:38:23 EST


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

--
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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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