LARGE COVALENT GOLD LABELS AND PROBES

一种新型的化学官能化，共价连接的金纳米颗粒，直径5 ~ 40nm，将被制备为胶体金的改进替代品。新颗粒将使用紧密排列的烷硫醇配体外壳来稳定：这些将被合成定制，以在稳定的颗粒上导入完全的水溶性和最大的生物相容性。粒径将由配体填充和配体与金的比例来控制，并研究不同的分离方法以达到尽可能窄的粒径范围。烷基链在相邻配体上的交联以包裹金核并增加稳定性也将被研究。修饰后的金标签将通过化学特异性反应基团与抗体和蛋白质进行共价交联，并结合到它们的配体中，也可以冻干用作标记试剂。标签属性将被调整，以消除非特异性结合，背景染色和聚集，这限制了胶体金的应用。新的偶联物将在哺乳动物细胞核的功能组织和小孢子虫的发育组织的研究中与等效胶体金探针并行进行评估。印迹测试将用于调整其特性，以便将来在快速诊断设备中使用。新型大尺寸金探针将取代胶体金探针，扩大其应用范围。共价偶联将使制备新的试剂具有重要的优势，在显微镜和结构生物学，这是不可行的胶体金。其卓越的设计和性能也将有助于开发更敏感和可靠的医疗诊断和新的设备格式。

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.