Directional conjugation of antibodies to nanoparticles for synthesis of multiplexed optical contrast agents with both delivery and targeting moieties
Directional conjugation of antibodies to nanoparticles for synthesis of multiplexed optical contrast agents with both delivery and targeting moieties
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DOI：
10.1038/nprot.2008.1
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发表时间：
2008-01-01
期刊：
影响因子：
14.8
通讯作者：
Sokolov, K.
中科院分区：
文献类型：
--
作者：
Kumar, S.;Aaron, J.;Sokolov, K.
分子光学成像技术在非侵入性和实时的亚细胞分辨率的分子生物标志物可视化方面显示出了希望。在这里，我们使用金纳米颗粒作为光学探针，在靶向生物标志物存在的情况下提供有意义的信号。我们提出了一种新的缀合技术来控制金纳米颗粒表面上的抗体的结合方向，以最大限度地提高抗体的功能。简而言之，异双功能接头，酰肼-聚乙二醇-二硫醇，用于将Fc或抗体的非结合区定向连接至金纳米颗粒表面。缀合策略允许在单个纳米颗粒上多路复用各种糖基化抗体。我们提出了一种方法来制备多功能纳米颗粒，通过将靶向和递送部分在同一纳米颗粒上，解决了细胞内生物标志物成像的挑战。整个方案的时间估计值类似于6 h。
Molecular optical imaging has shown promise in visualizing molecular biomarkers with subcellular resolution both noninvasively and in real-time. Here, we use gold nanoparticles as optical probes to provide meaningful signal in the presence of targeted biomarkers. We present a novel conjugation technique to control the binding orientation of antibodies on the surface of gold nanoparticles to maximize antibody functionality. Briefly, a heterobifunctional linker, hydrazide-polyethylene glycol-dithiol, is used to directionally attach the Fc, or nonbinding region of the antibody, to the gold nanoparticle surface. The conjugation strategy allows for multiplexing various glycosylated antibodies on a single nanoparticle. We present a method to prepare multifunctional nanoparticles by incorporating targeting and delivery moieties on the same nanoparticle that addresses the challenge of imaging intracellular biomarkers. The time estimate for the entire protocol is similar to 6 h.