This Small Business Innovation Research (SBIR) Phase II project will demonstrate the production of robust, versatile probes based on Bioluminescence Resonance Energy Transfer to Quantum Dots (BRET-Qdot®) technology. The main goal of this project is to develop a platform for target-directed in vivo imaging of tissue, such as whole tumors and organs, in animal models. To accomplish this goal, biotinylated targeting agents for the detection of important biomarkers, in combination with BRET-Qdot® probes with streptavidin functionality, will be developed to provide one-step target detection. The probes will represent the range of molecular sizes used for a majority of assays, from small molecules to antibodies, thus enabling detection of virtually any cell surface target molecule.
The broader impact/commercial potential of this project is the development of a sensitive probe reagent that will be broadly applicable to a wide range of preclinical assays including in vivo imaging of tumors, organs and various anatomical structures. In order to understand the molecular mechanisms underlying diseases, such as cancer, autoimmune diseases, and other diseases, there is increasing interest in advancing from cell-based assays to in vivo imaging of disease states in small animal models. The targeted BRET-Qdot® platform will provide a cost effective,convenient and effective alternative to competing fluorescence and bioluminescence imaging probe technology.
As a result of this award, Zymera has developed Bioluminescence Resonance Energy Transfer (BRET) probes, termed "BRET-Qdots," primarily for preclinical molecular imaging. BRET-Qdot probes have several copies of bioluminescent protein -- Luc8, a mutated Renilla luciferase -- covalently linked to the surface of a quantum dot. Coelenterazine-activated bioluminescence energy from the Luc8 luciferase is directly coupled to the quantum dot, which in turn emits light at wavelengths that are minimally absorbed or scattered in tissues, enabling visualization of molecular events inside living subjects. By combining the BRET-Qdot probes to agents targeting biological molecules such as cell-membrane receptors, the technology enabled imaging of early state metastases in animal models for pancreatic cancer. Other applications that were demonstrated as part of this project include ultra-sensitive cell-based assays for membrane receptors relevant to oncology research, and enzyme-linked immunoassays for antigen detection used in vaccine development. The broader impact of this project was the development and commercialization of a sensitive probe reagent that is broadly applicable to a wide range of preclinical assays including in vitro cell-based assays and in vivo imaging of tumors, organs and various anatomical structures. The technologies that were explored as part of this award enabled new capabilities applicable to bio-therapeutic research and to vaccine development.
