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; 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，保质期有限和成本导致免疫测定的非同位素标签的发展。替代填充方法包括酶免疫测定，化学发光测定法和荧光免疫测定法。时间分辨的荧光测定法实现了使用灯笼标签的测定法，与通过放射免疫测定所实现的标签相当。基于灯笼的生物分析的检测的主要缺点一直是水缓冲液中灯笼发光的淬火。通过在发光测量之前添加增强步骤，在商业时间分辨的氟免疫测定套件中已经解决了此问题。为了消除增强步骤并加快免疫测定的速度，提出了一类新的金属螯合剂。该多齿双功能螯合剂具有共价敏化剂的预期，将产生稳定且发光的灯笼标签。可以使用酶和标记的球蛋白进一步提高新标签的灵敏度。新的螯合剂还将有助于将各种标签（例如氧化还原，荧光和同位素）融合到各种生物分子上，以实现各种最终用途。除了免疫测定外，该技术还将开放新的研究领域，并导致在各种领域的应用，例如DNA杂交测定法，流程仪和时间分辨的荧光显微镜。拟议的商业应用：尽管在免疫测定中使用了非同位素标签，例如酶，化学发光和生物素，但通常需要提高灵敏度和特异性。提出的发光灯笼标签将允许使用时间分辨的荧光法以比市售套件更少的步骤进行敏感检测生物分析。其次，使用时间重新浏览的荧光测定法以比市售套件更少的步骤同时检测生物分析。其次，新试剂的同时检测不同的双重分析植物也很困难。除了荧光免疫测定外，其他潜在的应用包括流式细胞术，时间分辨荧光显微镜和原位DNA杂交。