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发展。替代的群体方法包括酶免疫测定法、化学发光测定法和荧光免疫测定法。时间分辨荧光法使得使用镧系元素标记的测定与通过放射免疫测定实现的测定相当。基于镧系元素的生物分析物检测的主要缺点是在水性缓冲液中镧系元素发光的猝灭。这个问题已经在商业时间分辨荧光免疫测定试剂盒中通过在发光测量之前添加增强步骤来解决。为了消除增强步骤并加快免疫测定，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，预计会得到稳定和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。新的螯合剂也将有助于纳入各种标签，例如，氧化还原、荧光和同位素，用于各种最终用途的各种生物分子。除了免疫分析，这项技术将开辟新的研究领域，并导致在各种领域的应用，如DNA杂交分析，流式细胞术，时间分辨荧光显微镜。拟定商业应用：虽然非同位素标记，如酶，荧光和生物素，用于免疫测定，提高灵敏度和特异性往往是可取的。所提出的发光镧系元素标记物将允许使用时间分辨荧光法在比市售试剂盒更少的步骤中灵敏地检测生物分析物。其次，使用时间旋转荧光法同时检测生物分析物，步骤比市售试剂盒少。第二，同时检测不同的双分析物，这是很难与其他技术，也将有可能与新试剂。除了荧光免疫测定，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。