Elucidating Protein/Ligand Recognition with Combined Surface Plasmon Resonance and Surface Enhanced Raman Spectroscopy.
Elucidating Protein/Ligand Recognition with Combined Surface Plasmon Resonance and Surface Enhanced Raman Spectroscopy.
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DOI：
10.1021/acs.analchem.7b04246
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发表时间：
2017-12-19
影响因子：
7.4
通讯作者：
Schultz ZD
中科院分区：
文献类型：
--
作者：
Kim JY;Zeng ZC;Xiao L;Schultz ZD
区分特异性和非特异性结合的能力对于评估蛋白质受体和配体之间的相互作用非常重要。表面等离子共振 (SPR) 光谱是一种测量结合事件的先进工具，但区分特异性和非特异性结合的能力仍然受到限制。为了解决这个问题，我们使用与表面增强拉曼散射 (SERS) 相关的 SPR 光谱。 SERS 光谱中存在的化学信息提供了对功能化纳米颗粒和蛋白质之间分子相互作用的深入了解，而这些相互作用是仅通过 SPR 无法检测到的。使用具有 Kretschmann 配置的定制仪器，我们成功演示了链霉亲和素功能化金纳米颗粒 (STV-NP) 在空气和流动磷酸盐缓冲盐水 (PBS) 中与固定在金膜上的生物素结合的同时亲和力和化学表征。 SPR 性能与之前的报告一致。观察到的链霉亲和素/生物素和 STV-NPs/生物素相互作用的关联常数 (KA)（分别为 2±1 ×107 M−1 和 2.4±0.3 ×1010 M−1）与文献值一致，并显示出与 STV-NPs 相关的强烈亲合力效应。 STV-NP 的 SERS 散射由表面等离子激元激发，并从安装在流体通道上的物镜收集。 SERS 光谱与 SPR 传感图同时记录，检测到的拉曼带提供了对结合事件的化学洞察。光谱的多变量曲线分辨率分析可以区分特异性结合和非特异性结合。这种无标记、实时且表面敏感的检测方法为蛋白质-配体结合亲和力测量提供化学信息。
The ability to distinguish between specific and non-specific binding is important for assessing the interactions between protein receptors and ligands. Surface plasmon resonance (SPR) spectroscopy is an advanced tool to measure the binding events, yet the ability to distinguish between specific and non-specific binding remains a limitation. To address this problem, we use SPR spectroscopy correlated with surface enhanced Raman scattering (SERS). The chemical information present in SERS spectra provides insight into the molecular interactions between functionalized nanoparticles and proteins, which are not detectable by SPR alone. Using a custom instrument with the Kretschmann configuration, we successfully demonstrate simultaneous affinity and the chemical characterization of streptavidin-functionalized gold nanoparticles (STV-NPs) binding to biotin immobilized on a gold film in both air and flowing phosphate buffered saline (PBS). The SPR performance is consistent with previous reports. The association constant (KA) for streptavidin/biotin and STV-NPs/biotin interactions observed (2±1 ×107 M−1 and 2.4±0.3 ×1010 M−1, respectively) agree with literature values and show a strong avidity effect associated with the STV-NPs. The SERS scattering from STV-NPs is excited by the surface plasmon polariton and collected from an objective lens mounted over the fluidic channel. The SERS spectra are recorded simultaneously with SPR sensorgram, and the detected Raman bands provide chemical insight into the binding event. Multivariate curve resolution analysis of the spectra can differentiate specific from non-specific binding. This label-free, real time and surface sensitive detection method provides chemical information to protein-ligand binding affinity measurements.
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影响因子：
7.4
作者：
Liang, Wenbin;Wang, Shaopeng;Festa, Fernanda;Wiktor, Peter;Wang, Wei;Magee, Mitchell;LaBaer, Joshua;Tao, Nongjian
通讯作者：
Tao, Nongjian
影响因子：
--
作者：
GIERGIEL, J;REED, CE;USHIODA, S
通讯作者：
USHIODA, S
影响因子：
7.4
作者：
McKee, Kristopher J.;Meyer, Matthew W.;Smith, Emily A.
通讯作者：
Smith, Emily A.
影响因子：
7.4
作者：
Nelson, BP;Grimsrud, TE;Corn, RM
通讯作者：
Corn, RM
影响因子：
3.3
作者：
Couture, Maxime;Zhao, Sandy Shuo;Masson, Jean-Francois
通讯作者：
Masson, Jean-Francois