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Versatile Bioconjugation Strategies of PEG-Modified Upconversion Nanoparticles for Bioanalytical Applications.
Biomacromolecules ( IF 5.5 ) Pub Date : 2020-05-11 , DOI: 10.1021/acs.biomac.0c00459 Uliana Kostiv 1 , Zdeněk Farka 2, 3 , Matthias J Mickert 4 , Hans H Gorris 4 , Nadiia Velychkivska 1 , Ognen Pop-Georgievski 1 , Matěj Pastucha 2, 3 , Eliška Odstrčilíková 2 , Petr Skládal 2, 3 , Daniel Horák 1
Biomacromolecules ( IF 5.5 ) Pub Date : 2020-05-11 , DOI: 10.1021/acs.biomac.0c00459 Uliana Kostiv 1 , Zdeněk Farka 2, 3 , Matthias J Mickert 4 , Hans H Gorris 4 , Nadiia Velychkivska 1 , Ognen Pop-Georgievski 1 , Matěj Pastucha 2, 3 , Eliška Odstrčilíková 2 , Petr Skládal 2, 3 , Daniel Horák 1
Affiliation
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Lanthanide-doped upconversion nanoparticles (UCNPs) display highly beneficial photophysical features for background-free bioimaging and bioanalysis; however, they are instable in high ionic strength buffers, have no functional groups, and are nonspecifically interacting. Here, we have prepared NIR-excitable UCNPs that are long-term colloidally stable in buffered media and possess functional groups. Heterobifunctional poly(ethylene glycol) (PEG) linkers bearing neridronate and alkyne or maleimide were attached to UCNPs via a ligand exchange. Streptavidin (SA)-conjugates were prepared by click reaction of UCNP@PEG-alkyne and SA-azide. Antihuman serum albumin pAbF antibody was modified with azide groups and conjugated to UCNP@PEG-alkyne via click reaction; alternatively, the antibody, after mild reduction of its disulfide bonds, was conjugated to UCNP@PEG-maleimide. We employed these nanoconjugates as labels for an upconversion-linked immunosorbent assay. SA-based labels achieved the lowest LOD of 0.17 ng/mL for the target albumin, which was superior compared to a fluorescence immunoassay (LOD 0.59 ng/mL) or an enzyme-linked immunoassay (LOD 0.56 ng/mL).
中文翻译：
用于生物分析应用的PEG修饰的上转换纳米粒子的通用生物共轭策略。
镧系元素掺杂的上转换纳米粒子（UCNPs）具有非常有益的光物理特性，可用于无背景的生物成像和生物分析。然而，它们在高离子强度缓冲液中不稳定，没有官能团，并且非特异性相互作用。在这里，我们准备了可在缓冲介质中长期胶体稳定并具有官能团的NIR可激发UCNP。带有神经氨酸酯和炔烃或马来酰亚胺的异双功能聚（乙二醇）（PEG）接头通过配体交换连接到UCNP。通过UCNP @ PEG-炔和SA-叠氮化物的点击反应制备链霉亲和素（SA）-缀合物。用叠氮基修饰抗人血清白蛋白pAbF抗体，并通过点击反应与UCNP @ PEG-炔偶联。或者，抗体在其二硫键温和还原后，将其与UCNP @ PEG-马来酰亚胺缀合。我们将这些纳米缀合物用作上转换连接的免疫吸附测定的标记。基于SA的标记物对目标白蛋白的最低LOD为0.17 ng / mL，优于荧光免疫测定法（LOD 0.59 ng / mL）或酶联免疫测定法（LOD 0.56 ng / mL）。
更新日期：2020-05-11
中文翻译：
用于生物分析应用的PEG修饰的上转换纳米粒子的通用生物共轭策略。
镧系元素掺杂的上转换纳米粒子（UCNPs）具有非常有益的光物理特性，可用于无背景的生物成像和生物分析。然而，它们在高离子强度缓冲液中不稳定，没有官能团，并且非特异性相互作用。在这里，我们准备了可在缓冲介质中长期胶体稳定并具有官能团的NIR可激发UCNP。带有神经氨酸酯和炔烃或马来酰亚胺的异双功能聚（乙二醇）（PEG）接头通过配体交换连接到UCNP。通过UCNP @ PEG-炔和SA-叠氮化物的点击反应制备链霉亲和素（SA）-缀合物。用叠氮基修饰抗人血清白蛋白pAbF抗体，并通过点击反应与UCNP @ PEG-炔偶联。或者，抗体在其二硫键温和还原后，将其与UCNP @ PEG-马来酰亚胺缀合。我们将这些纳米缀合物用作上转换连接的免疫吸附测定的标记。基于SA的标记物对目标白蛋白的最低LOD为0.17 ng / mL，优于荧光免疫测定法（LOD 0.59 ng / mL）或酶联免疫测定法（LOD 0.56 ng / mL）。
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