Novel multi-color, time-resolved, luminescent lanthanide reporters

描述（由申请人提供）：根据第一阶段SBIR，Lumiphore将使用其专有的镧系元素技术开发“下一代”多重报告染料，其物理特性上级于目前可用的报告染料。Lumiphore的镧系化合物具有比常规有机荧光材料更长的寿命、更大的斯托克斯位移和更尖锐的荧光峰轮廓，并且具有比市售的基于镧系的荧光产品更高的稳定性和亮度。在SBIR的努力下，Lumiphore提出开发用于多色时间分辨荧光的Lumi 4”- Tb（供体）。Lumi 4”-Tb最初由加州-伯克利大学开发，是一种基于二甲酰胺的大环螯合物，在标准水性缓冲液环境中稀释浓度下具有极高的量子产率（60%）、消光系数（26，000 M-1cm-1）和长寿命（2.7 ms）。Lumi 4 TM-Tb具有四个窄的发射峰，中心约为490 nm、545 nm、590 nm和620 nm。这些发射峰通过能量转移耦合到具有重叠激发最大值的宽范围的常规有机荧光团（受体）形成了所提出的双荧光系统的基础。我们还建议改变供体和受体之间的连接体特性，使得在其他方面相同的对的发射寿命是可辨别的。这将产生除了发射波长之外的附加的多路复用特性。拟议的研究将克服有机荧光染料的基本局限性，这些局限性限制了高通量筛选（HTS）药物发现，基因组筛选，快速DNA测序，临床诊断和荧光显微镜的荧光应用的灵敏度，速度和多路复用能力和可靠性。公共卫生相关性：拟议的研究将通过实现高度多重的诊断测定格式来影响公共卫生，该格式联合收割机快速周转时间和高测定通量效率。这些分析将用于高通量筛选（HTS）药物发现，基因组筛选，快速DNA测序，临床诊断和荧光显微镜应用的基础和应用生物医学研究。

DESCRIPTION (provided by applicant): Under this Phase I SBIR, Lumiphore will use its proprietary lanthanide technology to develop the `next-generation' of multiplex capable reporter dyes with photophysical properties superior to those currently available. Lumiphore's lanthanide compounds have longer life-times, a larger Stokes shift, and sharper fluorescent peak profiles that conventional organic fluorescent materials, and have greater stability and brightness than commercially available lanthanide- based fluorescence products. Under this SBIR effort, Lumiphore proposes to develop Lumi4"- Tb (Donor) for multi-color time resolved fluorescence. Lumi4"-Tb, originally developed at University of California - Berkeley, is an isophthalamide based macrocyclic chelate that has an exceptionally high quantum yield (60%), extinction coefficient (26,000 M-1cm-1), and long lifetime (2.7 ms) at dilute concentrations in standard aqueous buffer environments. Lumi4TM-Tb has four narrow emission peaks centered at approximately 490nm, 545nm, 590nm, and 620nm. These emission peaks form the basis of the proposed multicolor fluorescent system through energy transfer coupling to a wide range of conventional organic fluorophores (Acceptors) with overlapping excitation maxima. We propose to also vary the linker characteristics between Donor and Acceptors such that emission lifetimes of otherwise identical pairs are discernable. This will create an additional multiplex property in addition to emission wavelength. The proposed research will overcome fundamental limitations of organic fluorescent dyes that have limited the sensitivity, speed, and multiplexing capability and reliability of florescence applications for high throughput screening (HTS) drug discovery, genomic screening, rapid DNA sequencing, clinical diagnostics and fluorescence microscopy. PUBLIC HEALTH RELEVANCE: The proposed research will impact public health by enabling highly multiplexed diagnostic assay formats that combine fast turnaround times and high assay throughput efficiency. These assays will be used in high throughput screening (HTS) drug discovery, genomic screening, rapid DNA sequencing, clinical diagnostics and fluorescence microscopy applications for both basic and applied biomedical research.