purification of OKT3

Dr. Howard T. Petrie h-petrie at ski.mskcc.org
Wed Sep 4 17:07:34 EST 1996

In article <32218B1A.2643 at ulg.ac.be>, Nathalie Jacobs <N.Jacobs at ulg.ac.be>

> we try to purify the mouse antibody OKT3 (IgG2a) from the hybridoma 
> supernatant....We have several problems...

We have grown OKT3 in bioreactors with great success using Hybridoma
Serum-Free medium from GIBCO/Life Technologies.  The combination of
bioreactor production with serum-free medium generally results in
antibodies which come out of culture at 2-5mg/ml and >90% purity.  Final
cleanup is a simple DEAE column.  If you aren't inclined to buy a
bioreactor, it (H-SFM) works just as well in static culture, although the
concentrations (and relative purity) are less.  Still, it would have to be
better than 10% FBS.


Howard T. Petrie
Assistant member, Immunology Program
Memorial Sloan-Kettering Cancer Center
New York, NY U.S.A

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