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

描述(由申请人提供)：根据这一第一阶段SBIR，Lumiphore将使用其专有的镧系元素技术开发具有光物理性能优于现有产品的具有多重功能的“下一代”报告染料。Lumiphore的稀土化合物比传统的有机荧光材料具有更长的寿命、更大的斯托克斯位移和更清晰的荧光峰形，并且比商业上可用的基于镧系元素的荧光产品具有更高的稳定性和亮度。在这项SBIR工作中，Lumiphore建议开发用于多颜色时间分辨荧光的Lumi4“-Tb(供体)。Lumi4”-Tb最初由加州大学伯克利分校开发，是一种基于间苯二甲酰胺的大环络合物，在标准的水缓冲环境中具有极高的量子产率(60%)、消光系数(26,000 M-1 cm-1)和长寿命(2.7 ms)。Lumi4TM-Tb有四个窄的发射峰，分别位于大约490 nm、545 nm、590 nm和620 nm。这些发射峰构成了所提出的多色荧光体系的基础，通过与具有重叠激发峰的广泛的传统有机荧光团(受体)的能量转移耦合而形成。我们还建议改变供体和受体之间的连接子特性，以便可以辨别其他相同对的发射寿命。这将创建除了发射波长之外的额外的多路复用属性。拟议的研究将克服有机荧光染料的根本限制，这些限制限制了荧光应用于高通量筛选(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的灵敏度、速度、多重能力和可靠性。与公共健康相关：拟议的研究将通过实现结合快速周转时间和高分析吞吐量效率的高度多元化的诊断分析格式来影响公共健康。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，用于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.