Hi,
we try to purify the mouse antibody OKT3 (IgG2a) from the hybridoma
supernatant. We are using a home made sepharose A column (non HPLC).
We have several problems:
-a contamination with bovin IgG (the hybridoma is cultivated with RPMI
1640 + 20% SVF. Have somebody already try to cultivate them with
hybridoma medium without serum?)
-we don't obtain a lot of antibodies at the pH 4.5 (the pH for IgG2a).
We obtain higher concentration at pH 5.5(certainly bovin IgG)! We try
to concentrate the supernatant but everything have been detached at pH
5.5
I would appreciate any suggestion to increase the concentration of
purified OKT3.
Thank you in advance.
Nathalie Jacobs
