?detergent compatibility of mAb Immunoaffinity purification?

yuling luo yluo at pobox.upenn.edu
Wed Apr 19 09:58:53 EST 1995


Robert C. Colgrove (robin at alumni.caltech.edu) wrote:
: 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

I am having the same problem. I don't know if detergent will help or not 
but I added 0.1% cholic acid. If you have any success with your 
purification, please let me know.             yuling luo   upenn




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