IgG determination - (Sep/05/2001 )

I am producing momoclonal Ab's via a hollow fiber bioreactor. IgG1. The cells are viable and the conditions are right for proliferation. However, according to my co-worker there is little Ad produced < 0.1mg/ml after purification via protein A . I believe this to be false. I tried an Indirect Elisa, activity is there, but it is difficult to quantitate. HELP

If you have a mouse monoclonal antibody IgG1 you cannot use protein A for the purification of this antidody, because its low binding wiht this monoclonal antibody type. I recommend you protein L columns from PIERCE (20510). I've used it, and it works very well to purify monoclonal antibodies from mouse because the protein L does not bind Bovine Igs from de FCS.