The long-term goal of this project is the development of luminescent biomarkers containing lanthanide(III) complexes functionalized on macrocyclic ligands, "Quantum Dyes." Previous work has shown that functionalized europium(III) macrocyclic complexes are capable of covalent attachment to bio-substances and permit background free, time-gated luminescence measurements. However, for many practical applications their luminescence intensity had to be increased. The Phase I work resulted in: 1) The synthesis of markers containing multiple Eu-macrocycles, 2) Enhancement of the Eu-macrocycle luminescence by additives that act as energy-transfer donors or provide protection from quenching, and 3) An enhancer was found that promotes luminescence from the prototype non-functionalized terbium (III-macrocycle. An isomorphous pair of europium (III) and terbium (III) functionalized luminescent macrocycles will permit sensitive, precise comparative hybridization studies. In Phase II: 1) Sensitive tests for apoptosis and DNA synthesis will be developed, 2) The nonspecific binding of markers containing multiple Eu-macrocycles will be minimized, 3) Functionalized terbium(III) macrocycles will be synthesized and tested. 4) Preliminary work will be performed on how to maximize the luminescence from the combined use of europium (III) and terbium(III) macrocycles, and 5) Improvements to the instrumentation for detecting Quantum Dyes will be made.
