Rigid, specific, and discrete gold nanoparticle/antibody conjugates
Rigid, specific, and discrete gold nanoparticle/antibody conjugates
复制标题
DOI：
10.1021/ja0555668
复制
发表时间：
2006-03-01
影响因子：
15
通讯作者：
Kornberg, RD
中科院分区：
文献类型：
--
作者：
Ackerson, CJ;Jadzinsky, PD;Kornberg, RD
一个一般的方法，刚性，特定的标记与金簇的蛋白质已经设计。该方法依赖于谷胱甘肽单层保护金簇（MPC）与单链Fv抗体片段（scFv）的偶联，突变后呈现暴露的半胱氨酸残基。在MPC和scFv之间有效形成金-硫酸盐键取决于金簇的化学氧化活化。一旦形成，用还原剂处理MPC-scFv共轭物以淬灭簇反应性。该程序是用平均Au-71核的MPC和针对四聚体蛋白（流感神经氨酸酶）的scFv进行的。分离出MPC-scFv与神经氨酸酶缀合物，并通过冷冻电镜证实其存在4个金簇。
A general method of rigid, specific labeling of proteins with gold clusters has been devised. The method relies on the conjugation of a glutathione monolayer-protected gold cluster (MPC) with a single chain Fv antibody fragment (scFv), mutated to present an exposed cysteine residue. Efficient formation of a gold-thiolate bond between the MPC and scFv depends on activation of the gold cluster by chemical oxidation. Once formed, the MPC-scFv conjugate is treated with a reductant to quench cluster reactivity. The procedure has been performed with an MPC with an average Au-71 core and an scFv directed against a tetrameric protein, the influenza neuraminidase. A complex of the MPC-scFv conjugate with the neuraminidase was isolated, and the presence of four gold clusters was verified by cryoelectron microscopy.