title: Application of 4'-terpyridinylsulfanylethylamine resins for the purification of monoclonal antibodies by mixed-mode chromatography.
abstract: 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.
authors.affiliation: Centre for Green Chemistry, School of Chemistry, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Chunfang
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Centre for Green Chemistry, School of Chemistry, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Dale P
authors.initials: DP
authors.lastname: Fredericks
authors.affiliation: Centre for Green Chemistry, School of Chemistry, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Eva M
authors.initials: EM
authors.lastname: Campi
authors.affiliation: Centre for Green Chemistry, School of Chemistry, Monash University, Clayton, Victoria 3800, Australia. Electronic address: milton.hearn@monash.edu.
authors.forename: Milton T W
authors.initials: MT
authors.lastname: Hearn
mesh_terms: D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives* / Q000737:chemistry; D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D061067:Antibodies, Monoclonal, Humanized / Q000302:isolation & purification*; D016466:CHO Cells; D002853:Chromatography, Liquid / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000737:chemistry; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008024:Ligands; D011485:Protein Binding; D011725:Pyridines / Q000737:chemistry; D012117:Resins, Synthetic / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry
chemical_list: C000625959:4'-terpyridinylsulfanylethylamine; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D007074:Immunoglobulin G; D008024:Ligands; D011725:Pyridines; D012117:Resins, Synthetic; D013205:Staphylococcal Protein A; D015082:2,2'-Dipyridyl; D003543:Cysteamine; D012685:Sepharose; C023666:pyridine
keywords: Binding capacities; Binding sites; Mixed mode adsorbents; Monoclonal antibodies