LUMINESCENT LANTHANIDE CHELATES LABELS FOR IMMUNOASSAYS

用于免疫测定的发光镧系元素螯合物标签

• 批准号: 2865481
• 负责人: VISHWAS N JOSHI
• 金额: $ 10万
• 依托单位: NANOPROBES, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2865481
• 关键词: chelating agents; chemical synthesis; chromophore; fluorescent dye /probe; fluorimetry; high performance liquid chromatography; immunoconjugates; immunologic assay /test; luminescence; nuclear magnetic resonance spectroscopy; rare earth element; technology /technique development

Radiation hazards, limited shelf life, and costs have led to the development of non-isotopic labels for immunoassays. The alternate populate methods include enzyme immunoassay, chemiluminescence assay, and fluorimmunometric assay. Time-resolved fluorometry enables assays using lanthanide labels are comparable to those achieved by radioimmunoassays. The primary disadvantage of the lanthanide-based detection of bioanalytes has been the quenching of lanthanide luminescence in aqueous buffers. This problem has been addressed in commercial time-resolved fluoroimmunometric assay kits by adding an enhancement step before the luminescence measurement. To eliminate the enhancement step and speed up the immunoassays, a new class of metal chelator is proposed. This multidentate bifunctional chelating agent with covalently attached sensitizer is expected to give stable and luminescent lanthanide labels. The sensitivity of the new labels can further be improved using enzymes and labeled globulins. The new chelator will also help to incorporate a variety of labels, e.g., redox, fluorescent, and isotopic, onto various biomolecules for diverse end uses. Besides immunoassays, this technology will open new areas of research and lead to applications in a variety of fields, such as DNA hybridization assay, flow-cytometry, and time-resolved fluorescence microscopy. PROPOSED COMMERCIAL APPLICATION: Although non-isotopic labels, such as enzymes, chemiluminescent, and biotin, are used in immunoassays, improved sensitivity and specificity are often desirable. T he proposed luminescent lanthanide labels will permit sensitive detection of bioanalytes using time-resolved fluorometry in fewer steps than the commercially available kits. Secondly, simultaneous detection of bioanalytes using time-revolved fluorometry in fewer steps than the commercially available kits. Secondly, simultaneous detection of different bianalytes, which has been difficult with other techniques, will also be possible with the new reagents. Besides fluoroimmunometric assays, other potential applications include, flow cytometry, time-resolved fluorescence microscopy, and in situ DNA hybridization.

辐射危害、有限的保质期和成本导致了用于免疫测定的非同位素标记的开发。替代的填充方法包括酶免疫测定、化学发光测定和荧光免疫测定。时间分辨荧光测定法使使用镧系元素标记的测定与放射免疫测定所实现的测定相当。基于稀土元素的生物分析物检测的主要缺点是稀土元素发光在水性缓冲液中的猝灭。通过在发光测量之前添加增强步骤，该问题已在商业时间分辨荧光免疫测定试剂盒中得到解决。为了消除增强步骤并加速免疫测定，提出了一类新的金属螯合剂。这种多齿双功能螯合剂与共价连接的敏化剂有望产生稳定且发光的镧系元素标记。使用酶和标记球蛋白可以进一步提高新标记的灵敏度。新的螯合剂还将有助于将各种标记（例如氧化还原、荧光和同位素）结合到各种生物分子上，以实现不同的最终用途。除了免疫测定之外，该技术还将开辟新的研究领域，并在多种领域产生应用，例如 DNA 杂交测定、流式细胞术和时间分辨荧光显微镜。拟议的商业应用：尽管非同位素标记（例如酶、化学发光和生物素）用于免疫测定，但通常需要提高灵敏度和特异性。所提出的发光镧系元素标记将允许使用时间分辨荧光测定法以比市售试剂盒更少的步骤灵敏地检测生物分析物。其次，使用时间旋转荧光测定法同时检测生物分析物，其步骤比市售试剂盒更少。其次，使用新试剂也可以同时检测不同的双分析物，这在其他技术中是困难的。除了荧光免疫测定之外，其他潜在应用包括流式细胞术、时间分辨荧光显微镜和原位 DNA 杂交。