Many biological activities including cell division, cell death, immune response and intercellular communication depend on complex non-covalent interactions between DNA, proteins and other biomolecules. Fluorescence-based high throughput screening assays have largely been used to detect the activity of small molecule inhibitors for such interactions. While aforementioned screening methods have been widely successful, they are incompatible with naturally fluorescent molecules and heterogeneous solution mixtures. To address these limitations, we have worked to develop a new electrochemiluminescence (ECL) platform to investigate biomolecule binding and screen for therapeutic leads. Our ECL platform is based on the biomolecular interaction of the transcription factor binding domain DNA sequence GGG-ACT-TTCC and a luminophore conjugated NF-κB mutant. In developing this platform, we have immobilized DNA onto carboxylic acid terminated magnetic beads and characterized the modified electrode surface with XPS and TOF-SIMS. In parallel, we developed a strategy to synthesize a maleimide appended derivative of tris(bipyridine)ruthenium(II) (Ru(bpy)<sub>3</sub>) ECL emitter that can be conjugated to cysteine terminated NF-κB protein mutants. The ECL signal is generated upon binding between the immobilized DNA and luminophore tagged transcription factor.  Attenuation of this signal is observed with the incubation of hybridized DNA.<br/>

Development of electrochemiluminescent platforms to monitor biomolecule binding phenomena

256th National Meeting (2018)

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