EaStCHEM School of Chemistry, The University of Edinburgh, Edinburgh EH9 3FJ, U.K.
Centre for Inflammation Research, The University of Edinburgh, Edinburgh EH16 4UU, U.K.
Anal Chem. 2024 Nov 12;96(45):17978-17983. doi: 10.1021/acs.analchem.4c02860. Epub 2024 Oct 29.
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 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.
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相关。因此，在化学水平上监测 RTKs 的活性可以为评估药物疗效提供新的生物医学见解和方法。然而，直接监测激酶活性具有挑战性，并且最常用的方法依赖于 Western blot 和 ELISA 等技术。在此，我们报告了一种基于金纳米粒子的表面增强拉曼散射 (SERS) 传感器的开发，该传感器允许通过 Axl 酶实时监测报告肽 (Axltide) 的酪氨酸磷酸化。我们证明，我们的传感器具有很强的信号定位能力，并且我们能够通过化学磷酸化和酶促反应检测到报告肽的酪氨酸磷酸化，其峰变化与自发拉曼光谱中观察到的相似。通过监测 SERS 光谱，我们可以实时观察磷酸化的变化。
