LUMINESCENT LANTHANIDE CHELATES LABELS FOR IMMUNOASSAYS

辐射危害、有限的保质期和成本导致了用于免疫分析的非同位素标记的开发。可供选择的填充方法包括酶免疫法、化学发光法和荧光免疫法。时间分辨荧光法使使用稀土标记的分析可以与放射免疫分析相媲美。基于稀土元素的生物分析物检测的主要缺点是在水溶液中稀土元素的发光被猝灭。这个问题已经在商业时间分辨荧光免疫分析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，加快免疫分析的速度，提出了一种新的金属螯合剂。这种带有共价键合敏化剂的多齿双功能络合剂有望给出稳定和发光的稀土标记。使用酶和标记的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，如氧化还原、荧光和同位素，结合到各种生物分子上，用于不同的最终用途。除了免疫分析，这项技术还将开辟新的研究领域，并导致在不同领域的应用，如DNA杂交分析、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，如酶、化学发光和生物素，被用于免疫分析，但提高灵敏度和特异度往往是可取的。所提出的发光稀土标记将允许使用时间分辨荧光法以比商业上可用的试剂盒更少的步骤灵敏地检测生物分析物。其次，与市售试剂盒相比，使用时间旋转荧光法同时检测生物分析物的步骤更少。其次，使用新试剂也可以同时检测不同的生物分析物，这在其他技术中一直是困难的。除了荧光免疫分析外，其他潜在的应用包括流式细胞术、时间分辨荧光显微镜和DNA原位杂交。

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.