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The crystal structures of antibody 2D12.5 Fab bound to an yttrium-DOTA analogue and separately to a gadolinium-DOTA analogue is reported. The binding interactions between the antibody and metal-DOTA complex are indirectly affected by changing the metal, because there are no direct protein-metal interactions.

An antibody that binds rare earth complexes selectively is used as a docking station for a set of probe molecules, of particular interest for medical imaging and therapy. The monoclonal antibody 2D12.5, developed against the DOTA analogue Y-BAD conjugated to the immunogenic protein KLH is examined.

The synthesis, structure, and photophysical properties of several Tb(III) complexes with octadentate, macrotricyclic ligands that feature a bicapped topology and 2-hydroxyisophthalamide (IAM) chelating units are reported. These Tb(III) complexes exhibit highly efficient emission (Φtotal ≥ 50%), large extinction coefficients (ϵmax ≥ 20,000 M-1 cm-1), and long luminescence lifetimes (τH2O ≥ 2.45 ms) at dilute concentrations in standard biological buffers. The structure of the methyl-protected ligand was determined by single-crystal X-ray diffraction and confirms the macrotricyclic structure of the parent ligand; the amide groups of the methyl-protected cage compound generate an anion binding cavity that complexes a chloride anion. Once the ligand is deprotected, a conformational change generates a similar cavity, formed by the phenolate and ortho amide oxygen groups that strongly bind lanthanide ions. The Tb(III) complexes thus formed display long-term stability, with little if any change in their spectral properties (including lifetime, quantum yield, and emission spectrum) over time or in different chemical environments. Procedures to prepare functionalized derivatives with terminal amine, carboxylate, and N-hydroxysuccinimide groups suitable for derivatization and protein bioconjugation have also been developed. These bifunctional ligands have been covalently attached to a number of different proteins, and the terbium complexes' exceptional photophysical properties are retained. These compounds establish a new aqueous stability and quantum yield standard for long-lifetime lanthanide reporters. [ABSTRACT FROM AUTHOR]