mouse monoclonal purification

Andy Thompson a.s.thompson at qub.ac.uk
Tue Jan 23 12:07:35 EST 2001


I'm a bit of a newbie to antibody purification; learning on the hoof
(microbial ecology background). I need to purify 5 mouse cell lines in
RPMI medium. All are isotype IgG1. Now what affinity column is best to
use for maximun yield? Pharmacia and Pierce both reckon Protein is not
really suitable, and recommend Protein G or A/G. Sigma are otherwise,
and recommend the use of Protein A for mouse IgG1. Protein G has a high
affinity for bovine IgG, and since my medium contains foetal calf serum,
this might not be suitable. Pierce (and others) are pushing Protein L,
which seems to be a bit pricier; it has a high affinity for all mouse
IgG subclasses (the cell lines are clones, so not worried about mixing
other subclasses, because they shouldn't be there), but no affinity for
bovine IgG. is this the one to go for, or are there others I should
consider?
Please reply to me direct if at all possible.

thanks,
Dr. Andy Thompson
Research Fellow
Questor Centre
Queens University
Belfast
http://questor.qub.ac.uk
a.s.thompson at qub.ac.uk






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