LARGE COVALENT GOLD LABELS AND PROBES

将制备一种新型的化学功能化、共价连接的金纳米颗粒，直径为5-40 nm，作为胶体金的改进替代品。新粒子将使用紧密堆积的烷硫醇配体外壳进行稳定：这些将被人工定制，以引入完全的水溶性和稳定粒子的最大生物兼容性。颗粒大小将通过配体堆积和配体与金的比例来控制，并将研究不同的分离方法，以实现尽可能窄的尺寸范围。还将研究相邻配体上的烷基链的交联性，以包覆金核心并增加稳定性。修饰后的金标记物将通过化学上特定的反应基团结合到它们的配位配体中，以共价方式与抗体和蛋白质交联，并被冷冻干燥以用作标记剂。标记属性将进行调整，以消除限制胶体金应用的非特异性结合、背景染色和聚集。在哺乳动物细胞核的功能组织和小孢子体的发育组织的研究中，新的结合物将与同等的胶体金探针平行进行评估。印迹测试将被用来调整它们的特性，以便将来用于快速诊断设备。拟议的商业应用新的Isrge金探针将取代胶体金探针并扩大其应用范围。共价偶联将使制备新试剂具有重要的显微镜和结构生物学优势，而这是胶体金所不能实现的。其卓越的设计和性能还将使开发更灵敏和可靠的医疗诊断和新的设备格式成为可能。

A new type of chemically functionalized, covalently linked gold nanoparticles, 5 to 40 nm in diameter, will be prepared as improved replacements for colloidal gold. The new particles will be stabilized using a shell of closely-packed alkanethiol ligands: these will be synthetically tailored to import complete water-solubility and maximum biocompatibility upon the stabilized particles. Particle size will be controlled by ligand packing and the ratio of ligands to gold, and different separation methods will be investigated to achieve the narrowest possible size range. Cross-linking of the alkyl chains on adjacent ligands to encapsulate the gold core and increase stability will also be investigated. The modified gold labels will be covalently cross-linked to antibodies and proteins via chemically specific reactive groups incorporated into their coordinated ligands, and also lyophilized for use as labeling reagents. Label properties will be tuned to eliminate non-specific binding, background staining and aggregation which restrict the application of colloidal gold. The new conjugates will be evaluated in parallel with equivalent colloidal gold probes in studies of the functional organization of the mammalian cell nucleus, and the developmental organization of microsporida. Blot testing will be used to tune their properties for future use in rapid diagnostic devices. PROPOSED COMMERCIAL APPLICATION New lsrge gold probes will replace colloidal gold probes and extend their applications. Covalent conjugation will enable the preparation of new reagents with important advantages for microscopy and structural biology which are not feasible with colloidal gold. Their superior design and performance will also enable the development of more sensitive and reliable medical diagnostics and new device formats.