FAQs on Purification with Antibody Affinity Chromatography

I don’t know if Protein A or Protein L is best for my antibody purification. What should I try?

Protein A chromatography is the most frequently used affinity resins in biomanufacturing for capturing recombinant monoclonal antibodies. Protein A chromatography relies on the specific and reversible binding of an immobilized protein A ligand to antibodies, most notably Immunoglobulin G (IgG) through the heavy chain within the Fc region.

Protein L has a strong affinity for the variable region of an antibody’s kappa light chain. Protein L chromatography is therefore suitable for the purification of a wide range of antibody related molecules which do not bind to Protein A. Examples are antibody fragments (Fabs), domain antibodies (Dabs) and single chain variable fragments (scFv) as well as IgM and IgA.