LARGE COVALENT GOLD LABELS AND PROBES

一种新型的化学功能化，共价连接的金纳米粒子，直径5至40纳米，将被制备为胶体金的改进替代品。 新的颗粒将使用紧密堆积的烷乙氧基化配体的壳来稳定：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。 颗粒大小将通过配体填充和配体与金的比率来控制，并且将研究不同的分离方法以实现最大可能的尺寸范围。 还将研究相邻配体上的烷基链的交联以包封金核心并增加稳定性。 修饰的金标记物将通过结合到其配位体中的化学特异性反应基团与抗体和蛋白质共价交联，并且也被冻干以用作标记试剂。通过调整标记物的性质，可以消除限制胶体金应用的非特异性结合、背景染色和聚集现象。 新的共轭物将在平行评价等效的胶体金探针的哺乳动物细胞核的功能组织的研究，和小孢子虫的发育组织。印迹测试将用于调整它们的性质，以便将来在快速诊断设备中使用。建议的商业应用新lsrge金探针将取代胶体金探针，并扩大其应用。 共价结合将使新试剂的制备具有重要的优势，显微镜和结构生物学，这是不可行的胶体金。 它们的上级设计和性能也将有助于开发更灵敏、更可靠的医疗诊断和新的设备形式。

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.