purification ployclonal Ab

Tina anfortin at webnet.qc.ca
Thu Nov 2 22:46:34 EST 2000

I purified a rabbit polyclonal antibody on an NHS-activated sepharose
mini-column. I got approx. 2.5 mg of IgG from 5 mL of starting serum. After
this I put this antibody solution on dialysis O/N. At this step all things
seems OK. I also concentrated the antibody solution by centrifugation on
CENTRICON membrane mounted tube. Surprise .... the antibody seems to have
precipitated (many particles in suspension) after the Centricon step !!!!
What's happens ? Is it possible to recover a clear solution and the
antibodies? I took OD 280 and it gives a greatly decreased antibody quantity
versus the quantity before dialysis !!!

I would appreciate some help or ideas because it is the first time I've got
this result !
Thanks a lot !

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