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Near Infrared Probes for Biochemical, Cellular, and Whole Animal Analysis of Disease Processes

机译:近红外探头用于生化,细胞和疾病过程的全动物分析

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The study of disease processes requires a number of tools for detection of proteins and biomarkers in cell and animal based assays. Near infrared (NIR) technologies offer the advantage of high signal without interference from background producing factors such as tissues, blood, or plastics. NIR fluorescence quenching biochemical assays employing a novel NIR quencher are homogeneous and sensitive. NIR-based immunocytochemical assays offer a means of quantitatively evaluating cell signaling pathways. The technology can be extended to the development of targeted molecular imaging agents for disease analysis in animal models. We describe here model assays for each of these categories. A fluorescence quenching caspase-3 assay was developed employing a novel, broadly applicable quencher dye suitable for use with both visible and NIR dye chemistries. An NIR cell based assay is described for assessment of phosphorylation of p53 in response to a cellular stimulus. Finally, we describe the development and application of a targeted NIR optical imaging agent for monitoring tumor growth in whole animals. The NIR biochemical and cell based assays are robust with Z' factors greater than 0.7. The use of an IRDye800CW-labeled cyclic RGD peptide is presented as a model for development and application of targeted imaging agents. NIR technologies are compatible with the complete spectrum of assay needs for disease analysis and therapeutic development.
机译:疾病方法的研究需要许多工具用于检测细胞和动物的测定中的蛋白质和生物标志物。近红外线(NIR)技术提供高信号的优势,而不会受到组织,血液或塑料等背景产生因子的干扰。使用新型NIR猝灭剂的NIR荧光猝灭生物化学测定是均匀和敏感的。基于NIR的免疫细胞化学测定提供了定量评估细胞信号传导途径的方法。该技术可以扩展到动物模型中患者疾病分析的靶分子成像剂的发展。我们在这里描述了每个类别的模型测定。开发了一种荧光猝灭的Caspase-3测定,采用新颖的广泛适用的猝灭剂染料,适用于可见和NIR染料化学物质。描述基于Nir细胞基的测定用于评估p53的磷酸化响应细胞刺激。最后,我们描述了针对靶向NIR光学成像剂的开发和应用,用于监测全动物的肿瘤生长。 NIR生物化学和基于细胞的测定具有大于0.7的Z'因子的稳健。使用IRDYE800CW标记的环状RGD肽作为靶向成像剂的开发和应用的模型。 NIR Technologies与疾病分析和治疗发展的完整测定谱兼容。

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