Upconverting Chelates for Cancer Detection and Diagnosis

这项拟议工作的长期目标是通过强大的新分子上转换标记来增强诊断、预测和治疗癌症的工具。这些标记具有许多潜在的用途，包括体外诊断、体内疾病组织的检测和标记，以及在微流控系统中进行高通量筛选。这项拟议工作的近期目标是通过应用于组织样本的体外免疫诊断，更好地了解和开发我们的分子上转换子的能力。这种新型的革命性的报道是基于稀土(稀土)络合物的上转换。这些材料没有自发荧光背景，不进行光漂白，使用红外激发(光毒性较小，提供良好的组织穿透性，可以使用二极管激光提供)，并且具有窄的发射波段(这有利于多路分析)。上转换络合物比上转换荧光粉(也是在SRI开发的)要小得多；上转换络合物的分子量为几百。我们将把上转换络合物应用于组织样本的免疫诊断，其中多路传输。光漂白的能力和缺失将是非常有价值的。在第一阶段的工作(R21)中，我们的具体目标包括生产一种上转换螯合物并测量其光学性质，组装上转换显微镜以供其使用，将螯合物与抗体结合，并在组织细胞系上测试螯合抗体探针。在第二阶段(R33)，我们将把螯合物扩展到多重分析，具体目标是准备两个额外的上转换螯合物，扩展上转换显微镜以检测所有三个螯合物，并同时检测淋巴瘤组织样本中的三个抗原。这项工作将结合光学物理学家、化学家、免疫学家、癌症生物学家和病理学家的努力。

The long term objectives of this proposed work are to enhance the tools for diagnosing, predicting, and treating cancer through powerful new molecular upconverting labels. These labels have a number of potential uses, including in vitro diagnostics, in vivo detection and demarkation of diseased tissue, and incorporation in microfluidic systems for high throughput screening. The near term objective of this proposed work is to better understand and develop the capabilities of our molecular upconverters through application to in vitro immunodiagnosis of tissue samples. This new and revolutionary type of reporter is based on upconversion in lanthanide (rare earth) chelates. These materials have no autofluorescent background, do not photobleach, use infrared excitation (which is less phototoxic, offers good tissue penetration, and can be provided using diode lasers), and have narrow emission bands (which is advantageous for multiplexed assays). The upconverting chelates are much smaller than the upconverting phosphors (also developed at SRI); the upconverting chelates have molecular weights of a few hundred. We will apply upconverting chelates to immunodiagnosis of tissue samples, where the multiplexing. capabilities and absence of photobleaching will be of great value. In the first phase of work (R21), our specific aims include producing one type of upconverting chelate and measuring its optical properties, assembling an upconverting microscope for its use, conjugating the chelate to antibodies, and testing the chelate-antibody probe in assays on tissue cell lines. In the second phase (R33), we will extend the chelates to multiplexed assays, with specific aims of preparing two additional upconverting chelates, extending the upconverting microscope to detect all three chelates, and simultaneously detecting three antigens in lymphoma tissue samples. This work will combine the efforts of optical physicists, chemists, immunologists, a cancer biologist, and a pathologist.