Ailsa Geddis, Lorena Mendive-Tapia, Audreylia Sujantho, Erica Liu, Sarah McAughtrie, Richard Goodwin, Marc Vendrell and Colin J. Campbell*,
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引用次数: 0
摘要
受体酪氨酸激酶(RTKs)的失调已被证明与癌细胞增殖和耐药性有关。因此,在化学水平上监测 RTKs 的活性可以提供新的生物医学见解和评估药物疗效的方法。然而,直接监测激酶活性具有挑战性,最常见的是依赖 Western 印迹法和 ELISAs 等体外技术。在此,我们报告了基于金纳米粒子的表面增强拉曼散射(SERS)传感器的开发情况,该传感器可实时监测 Axl 酶对报告肽(Axltide)的酪氨酸磷酸化。我们的研究表明,我们的传感器具有很强的信号定位能力,能够通过化学磷酸化和酶法检测报告肽的酪氨酸磷酸化,其峰值变化与自发拉曼光谱中观察到的峰值变化相似。通过监测 SERS 光谱,我们可以实时观察磷酸化的变化。
Label-Free SERS Sensors for Real-Time Monitoring of Tyrosine Phosphorylation
Dysregulation of receptor tyrosine kinases (RTKs) has been shown to correlate with cancer cell proliferation and drug resistance. Thus, monitoring the activity of RTKs at a chemical level could provide new biomedical insights and methods to assess the drug efficacy. However, direct monitoring of kinase activity is challenging and most commonly relies on in vitro techniques such as Western blotting and ELISAs. Herein, we report the development of a gold nanoparticle-based surface-enhanced Raman scattering (SERS) sensor, which allows the real-time monitoring of tyrosine phosphorylation of a reporter peptide (Axltide) by the Axl enzyme. We demonstrate that our sensor shows strong signal localization, and we are able to detect tyrosine phosphorylation of the reporter peptide through chemical phosphorylation and enzymatically with similar peak changes to those observed in the spontaneous Raman spectra. Through monitoring the SERS spectrum, we can observe changes in phosphorylation in real time.
期刊介绍:
Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.