SBIR Phase II: In vivo Fluorescence Imaging Kit for Cell Proliferation.

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This Small Business Innovation Research (SBIR) Phase II project proposes to develop an in vivo fluorescent imaging kit for cell proliferation that addresses the need to study cell growth processes in real time. Proof-of-concept has been demonstrated for a novel method of imaging DNA replication in vivo that is non-toxic and does not interfere with cellular metabolic processes. The method utilizes the unique specificity and high rate of reaction of bio-orthogonal ligation chemistry, and it has shown that in vivo labeling of DNA molecules with this novel bio-orthogonal mechanism yields fast, precise labeling of cell proliferation of cells in their natural environments. Most importantly, the method is minimally invasive, results in reliable incorporation of both the nucleotides and the label, and does not require cell lysis, DNA strand separations, or any of the abrasive treatments characteristic of cell proliferation assays currently on the market. The company plans to develop new methods for the usage of its technology with Fluorescently Activated Cell Sorting (FACS) protocols and testing different formats, such as tissues and different cell lines, for the applicability of this method.The broader impact/commercial potential of this project, if successful, is a Fluorescent Imaging Kit that can be routinely used with such advanced techniques as high content screening, high throughput screening for drug-discovery, and ADME-TOX assays, as well as with more traditional cell biology and molecular biology settings. The unique advantage of the proposed product is that it allows cell monitoring over prolonged time periods, up to several days. The product will have applications in multiple scientific disciplines, from cancer biology to stem cell biology, and streamline experimental protocols. The proposed assays are especially tailored to be fully compatible with high content screening, one of the major technologies at the forefront of personalized medicine, most notably in the field of oncology, and as such it will directly contribute to the advancement of new therapies, better diagnostics, and more efficient treatment plans. The method also will allow cutting costs of reagents by eliminating the need to repeat experiments multiple times to monitor different markers, and by streamlining research aspects of early stage drug discovery.