AMPLIFIED FLUORESCENT REPORTERS FOR AVIDIN-BIOTIN TESTS

这项拟议研究的长期目标包括制备带有放大荧光标记的亲和素、链霉亲和素和生物素衍生物，用于各种亲和素-生物素技术应用，包括免疫学、鱼类诊断和流式细胞术。所提出的报告放大共轭将增加信噪比，增强灵敏度，提高检测分辨率。使用原型扩增标签的初步结果表明，它们具有提供信号放大的潜力。本研究项目致力于改进标记的结构以进一步提高其适用性，具体包括以下目标：i)合成和评价12个新的放大荧光标记：2)标记与亲和素、修饰的亲和素和链霉亲和素的偶联，3)标记蛋白质及其生物素结合的荧光研究，4)扩增蛋白结合物与商业试剂在标准FISH分析中的荧光显微镜检测的比较，以及5)扩增标记生物素衍生物的合成和评价作为亲和素-生物素络合物的检测试剂。关于扩增的偶联物的结构要求的假设将通过与商业产品的直接比较来验证。该项目的健康相关性是基于放大的记者在被广泛接受的诊断方案中的应用。未来的发展将特别针对在流式细胞术和共振能量转移研究中的商业应用。建议的商业应用：放大的荧光探针在生物研究和医疗诊断方面有相当大的商业潜力。

The long-term objectives of the proposed research include preparation of avidin, streptavidin, and biotin derivatives labeled with amplified fluorescent tags for use in a variety of avidin-biotin technology applications, including immunology, FISH diagnostics, and flow cytometry. The proposed reporter amplified conjugates will increase signal-to-noise ratio, enhance sensitivity, and improve resolution of detection. Preliminary results using prototype amplified tags demonstrated their potential to provide signal amplification. This research project focuses on modification of the labels' structures to further enhance their suitability and includes the following specific goals: I) synthesis and evaluation of 12 novel amplified fluorescent labels: 2) conjugation of the labels with avidin, modified avidin, and streptavidin, 3) fluorimetric study of labeled proteins and their biotin binding, 4) comparison of amplified protein conjugates to commercial reagents in standard FISH analysis with fluorescent microscopy detection, and 5) synthesis and evaluation of amplified-labeled biotin derivatives as detection reagents for avidin-biotin complexes. The hypothesis concerning structural requirements of the amplified conjugates will be validated by direct comparison to commercial products. The health-relatedness of the project is based on application of the amplified reporters in well accepted diagnostic protocols. Future development will address specifically commercial applications in flow cytometry and resonance energy transfer studies. PROPOSED COMMERCIAL APPLICATIONS: There is a considerable commercial potential for the amplified fluorescence probes in biological research and medical diagnostics.