Purification of antibodies using the synthetic affinity ligand absorbent mabsorbent a2p

Abstract

A protocol for the purification of polyclonal antibodies from ovine serum using the synthetic protein A absorbent MAbsorbent A2P is described. Clarified serum is loaded directly onto the affinity column without prior adjustment and albumin and unwanted serum components are washed from the column using a sodium octanoate buffer before elution of bound antibodies. MAbsorbent A2P was shown to bind -27 mg ml-1 of polyclonal immunoglobulin under overloading conditions, with eluted IgG purities of >90% and minor levels of albumin (-1%). The anticipated time required to complete the purification protocol is 6-7 h. Although the protocol is similar to methods utilized for antibody purification using chromatography with protein A derived from the cell wall of the microorganism Staphylococcus aureus or protein G from Streptococcus as the affinity ligands, affinity absorbents based on synthetic ligands offer a number of advantages to compounds derived from biological sources, in particular robustness, relatively low cost, ease of sanitization and, in principle, lack of biological contamination. © 2007 Nature Publishing Group.