Application of 4′-terpyridinylsulfanylethylamine resins for the purification of monoclonal antibodies by mixed-mode chromatography

• Use of a new class of mixed mode adsorbents for mAb purification documented.
• The performance of this adsorbent compared to a Protein-A based affinity resin.
• Key attributes of selectivity, binding capacity and recyclability demonstrated.
• Preferred binding site of a humanised IgG1 mAb shown using papain digest fragments.

In this study, a pyridine-based compound, 4′-terpyridinylsulfanylethylamine (4′-TerPSEA), has been employed as a ligand to purify via mixed-mode chromatographic procedures a humanised monoclonal antibody of the IgG1 sub-class directly from crude supernatants derived from cultured CHO cells. The antibody binding capacity, selectivity and reusability of the adsorbent, derived from the immobilisation of this ligand onto Sepharose FF™, were compared to a Protein A affinity resin. The chromatographic performance of this mixed mode adsorbent was similar to that shown by the Protein A-based adsorbent with this IgG1 mAb. In addition, the IgG1 mAb was able to bind to the immobilised 4′-TerPSEA under reducing conditions. Through the use of papain-digested IgG1 mAb, fractionated with both the 4′-TerPSEA and Protein A adsorbents, it was found that this IgG1 mAb preferentially bound to the immobilised 4′-TerPSEA Sepharose FF™ resin through its Fc region.

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