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

这个小型企业创新研究(SBIR)第二阶段项目建议开发一种用于细胞增殖的体内荧光成像试剂盒，以满足实时研究细胞生长过程的需求。概念验证已经证明，一种新的方法，成像DNA复制在体内，是无毒的，不干扰细胞的代谢过程。该方法利用了生物正交连接化学独特的特异性和高反应速度，并表明用这种新的生物正交机制在体内标记DNA分子可以快速、准确地标记细胞在其自然环境中的细胞增殖。最重要的是，这种方法是微创的，可以可靠地结合核苷酸和标记，不需要细胞裂解、DNA链分离或任何目前市场上细胞增殖分析特有的研磨处理。该公司计划开发将其技术与荧光激活细胞分选(FACS)协议一起使用的新方法，并测试不同的格式，如组织和不同细胞系，以确保该方法的适用性。如果该项目成功，该项目的更广泛的影响/商业潜力是荧光成像试剂盒，它可以常规用于高含量筛选、用于药物发现的高通量筛选和ADME-TOX分析等先进技术，以及更传统的细胞生物学和分子生物学设置。拟议产品的独特优势是，它允许对细胞进行长时间的监测，最长可达几天。该产品将在从癌症生物学到干细胞生物学的多个科学学科中得到应用，并简化实验方案。建议的分析特别是为了与高含量筛查完全兼容而量身定做，高含量筛查是个性化医学的前沿主要技术之一，尤其是在肿瘤学领域，因此它将直接有助于新疗法的进步、更好的诊断和更有效的治疗计划。该方法还将通过消除多次重复实验来监测不同的标记物，并通过简化早期药物发现的研究方面，来削减试剂的成本。

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.