IgG purification

Mike Preston preston at warren.med.harvard.edu
Thu Jun 1 07:43:05 EST 1995


In article <852 at kiwi.ukc.ac.uk>, mrr1 at ukc.ac.uk (M.R.Reece-Ford) wrote:

>   I have a CHO cell line that produces a chimeric IgG4 that unfortunately has
> to be cultured in serum containing media (8% feotal calf serum). This FCS also
> contains bovine IgG which I would like to selectively remove from the culture
> supernatant. Has anyone had any experience with this problem? I have read that
> Pharmacia's Protein G Fast Flow 4 will remove the bovine IgG has anyone tried
> it? Finally how about purifying the FCS before addition to the media, is this
> feasible or would maintaining sterility of the serum be to difficult (the FCS
> has components that would be denatured if autoclaved). Thanks in advance.
> 
> Matthew

Matthew,
You are correct, you can preclear your fetal calf serum of bovine IgG by
running it over a protein G affinity column prior to adding it to your
media.  You will obviously have to filter sterilize your media after adding
the FCS.  The other alternative is to wean your CHO cells to a serum free
media.  Gibco has a serum free media for CHO cells called CHO-S-SFM II. 
Although it is formulated for growth of CHO cells in suspension in spinner
flasks (it contains a surfactant), I routinely grow them in regular tissue
culture flasks.  The cells tend to clump but they grow fine and and produce
comparable levels of chimeric antibodies. Gibco tech service will probably
tell you that this media is not optimized for growing CHO cells in regular
flasks.  It is probably not but I have found that it works just fine for
producing chimeric antibodies.
> --
Good luck,
Mike Preston
Channing Laboratory
Brigham and Women's Hospital and Harvard Medical School
180 Longwood Ave. 
Boston, MA 12115
preston at warren.med.harvard.edu
617-432-0152



More information about the Immuno mailing list