Synthesis of substituted terpyridine ligands for use in protein purification

A number of 4,4″-disubstituted terpyridines bearing a 4′-thioethanamine linker, and regioisomers thereof, have been synthesised from 4-substituted picolinates. The substituted terpyridines were immobilised onto epoxy-activated Sepharose™ FF gel, creating functionalised solid supports for the fractionation of proteins—including antibodies—by mixed mode affinity chromatography. The metal chelating properties of the immobilised ligand render the stationary phase also amenable for use in immobilised metal-ion affinity chromatography (IMAC), and have been demonstrated with the purification of suitably tagged green fluorescent protein (GFP). [Display omitted]