?detergent compatibility of mAb Immunoaffinity purification?

Robert C. Colgrove robin at alumni.caltech.edu
Tue Apr 18 14:52:00 EST 1995


Hi
I'm making recombinant viral envelope proteins in baculo
and purifying with a monoclonal immunoaffinity column I made.
This works great except that the protein goes into the column
dilute and impure but soluble (by western on gel filtration fractions)
and comes off pure concentrated and aggregated.
I am playing around with different kinds of elutions 'n stuff
but am also wondering about  loading, washing and eluting all in the
presence of mild detergent. As a virologist, I'm a little weak on the 
antibody biochemistry and I have been unable to find a straight answer
about which detergents would be least likely to torch the column
and/or the antigen/antibody complex. I know I do western washes with tween
so it can't be completely lethal.  Reading the detergent refs I can
find, I am trying Triton x-100, CHAPS, and octylglucoside
but it occurred to me to petition this august assembly for the collective
wisdom of  the net.group. Any thoughts?
many thanks,
			robin colgrove
			division of infectious diseases
			harvard school of medicine






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