Antibodies Irreversibly Adsorb to Gold Nanoparticles and Resist Displacement by Common Blood Proteins
Antibodies Irreversibly Adsorb to Gold Nanoparticles and Resist Displacement by Common Blood Proteins
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DOI：
10.1021/acs.langmuir.9b01900
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发表时间：
2019-08-13
期刊：
影响因子：
3.9
通讯作者：
Driskell, Jeremy D.
中科院分区：
文献类型：
--
作者：
Ruiz, Guadalupe;Ryan, Nicki;Driskell, Jeremy D.
金纳米颗粒(AuNPs)具有蛋白质功能化的表面特性，已被开发用于许多纳米生物技术的应用。蛋白质和纳米颗粒之间的强相互作用对于形成稳定的结合物以实现这些新兴技术的潜力至关重要。在这项工作中，我们检查了吸附在金纳米颗粒上的蛋白质层在可能导致纳米颗粒表面蛋白质交换的生理环境压力下的稳健性。通过纳米粒子跟踪分析，研究了常见血浆蛋白(转铁蛋白、人血清白蛋白、纤维蛋白原)和抗辣根过氧化物酶抗体在AuNPs上的吸附作用。我们的数据表明，每个蛋白质在饱和时都形成了一层蛋白质，并且结合物相对于AuNP核心的最大尺寸增加与蛋白质尺寸有关。每种蛋白质与AuNP的结合亲和力是从Hill方程的吸附等温线的最佳拟合中提取的。该抗体表现出最大的亲和力(K-d=15.2+/-0.8 nm)，接近于其他血浆蛋白亲和力的20-65倍。制备抗体-AuNP偶联物，纯化后悬浮于血浆蛋白溶液中，评价抗体层的稳定性。酶介导的检测证实，抗体-AuNP的相互作用是不可逆的，吸附的抗体抵抗血浆蛋白的置换。这项工作提供了对抗体-AuNP使能技术在生物系统中的能力和潜在限制的洞察。
Gold nanoparticles (AuNPs) functionalized with proteins to impart desirable surface properties have been developed for many nanobiotechnology applications. A strong interaction between the protein and nanoparticle is critical to the formation of a stable conjugate to realize the potential of these emerging technologies. In this work, we examine the robustness of a protein layer adsorbed onto gold nanoparticles while under the stress of a physiological environment that could potentially lead to protein exchange on the nanoparticle surface. The adsorption interaction of common blood plasma proteins (transferrin, human serum albumin, and fibrinogen) and anti-horseradish peroxidase antibody onto AuNPs is investigated by nanoparticle tracking analysis. Our data show that a monolayer of protein is formed at saturation for each protein, and the maximum size increase for the conjugate, relative to the AuNP core, correlates with the protein size. The binding affinity of each protein to the AuNP is extracted from a best fit of the adsorption isotherm to the Hill equation. The antibody displays the greatest affinity (K-d = 15.2 +/- 0.8 nM) that is similar to 20-65 times stronger than the affinity of the other plasma proteins. Antibody-AuNP conjugates were prepared, purified, and suspended in solutions of blood plasma proteins to evaluate the stability of the antibody layer. An enzyme-mediated assay confirms that the antibody- AuNP interaction is irreversible, and the adsorbed antibody resists displacement by the plasma proteins. This work provides insight into the capabilities and potential limitations of antibody-AuNP-enabled technologies in biological systems.