As a service core, we purifiy MAbs all the time by FPLC. We have purified
IgG from serum, ascites, and conditioned media. We apply the sample to
Protein G or rProtein A columns (1 ml or 10 ml depending on capacity -
Pharmacia) and elute the IgG with low pH glycine or citrate buffer,
neutralizing with 1M Tris or Phosphate. A buffer-exchange step is usually
necessary, but the IgG is >95% pure by SDS-PAGE. We always tell our clients
to check the flow thru in case a particular subclass of IgG doesn't bind,
but so far this hasn't appeared to be a problem.
Cris
Cristine Charlesworth, Ph.D.
Mayo Protein Core Facility
Mayo Foundation
Tel: 507 284-0912
Fax: 507 284-2053
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