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

摘要

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 IgG(1) 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 (TM), 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 IgG(1) mAb. In addition, the IgG(1) mAb was able to bind to the immobilised 4'-TerPSEA under reducing conditions. Through the use of papain-digested IgG(1) mAb, fractionated with both the 4'-TerPSEA and Protein A adsorbents, it was found that this IgG(1) mAb preferentially bound to the immobilised 4'-TerPSEA Sepharose FF (TM) resin through its F-c region. (C) 2014 Elsevier Inc. All rights reserved.