Centre for Personalized Nanomedicine, Australian Institute for Bioengineering and Nanotechnology , The University of Queensland , Brisbane , Queensland 4072 , Australia.
Centre for Advanced Imaging, Australian Institute for Bioengineering and Nanotechnology , The University of Queensland , Brisbane , Queensland 4072 , Australia.
Anal Chem. 2018 Sep 4;90(17):10377-10384. doi: 10.1021/acs.analchem.8b02216. Epub 2018 Aug 20.
Highly sensitive, multiplexed detection of soluble cancer protein biomarkers can facilitate early cancer screening as well as enable real-time monitoring of patients' sensitivity and resistance to therapy. Current technologies for detection of soluble cancer protein biomarkers, e.g., enzyme-linked immunosorbent assay, however, suffer from limited sensitivity, as well as the requirement of expensive monoclonal antibodies, which undergo the quality variability. Herein, we propose a sensitive, cheap, and robust surface-enhanced Raman scattering technology to detect a panel of soluble cancer protein biomarkers, including soluble programmed death 1 (sPD-1), soluble programmed death-ligand 1 (sPD-L1) and soluble epithermal growth factor receptor (sEGFR), which are related to disease progression and treatment efficacy. In this assay, gold-silver alloy nanoboxes that have strong Raman signal enhancement capability were used as plasmonic nanostructures to facilitate highly sensitive detection. In addition, nanoyeast single-chain variable fragments were utilized as mAb alternatives to allow specific and stable protein capture performance. We successfully detected sPD-1, sPD-L1, and sEGFR with a limit of detection of 6.17 pg/mL, 0.68 pg/mL, and 69.86 pg/mL, respectively. We further tested the detection of these three soluble cancer protein biomarkers in human serum and achieved recovery rates between 82.99% and 101.67%. We believe our novel platform that achieves sensitive, multiplexed, and specific detection of soluble cancer protein biomarkers could greatly benefit cancer treatment and improve patient outcome.
高灵敏度、多重检测可溶性癌症蛋白生物标志物可促进早期癌症筛查，并能够实时监测患者对治疗的敏感性和耐药性。然而，目前用于检测可溶性癌症蛋白生物标志物的技术，如酶联免疫吸附测定法，存在灵敏度有限的问题，而且需要昂贵的单克隆抗体，这些抗体存在质量变异。在此，我们提出了一种灵敏、廉价、稳健的表面增强拉曼散射技术，用于检测一组可溶性癌症蛋白生物标志物，包括可溶性程序性死亡受体 1（sPD-1）、可溶性程序性死亡配体 1（sPD-L1）和可溶性表皮生长因子受体（sEGFR），这些标志物与疾病进展和治疗效果相关。在该检测方法中，具有强拉曼信号增强能力的金银合金纳米盒被用作等离子体纳米结构，以实现高灵敏度检测。此外，纳米酵母单链可变片段被用作 mAb 的替代品，以实现特异性和稳定的蛋白捕获性能。我们成功地检测到 sPD-1、sPD-L1 和 sEGFR 的检测限分别为 6.17pg/mL、0.68pg/mL 和 69.86pg/mL。我们进一步在人血清中测试了这三种可溶性癌症蛋白生物标志物的检测，回收率在 82.99%至 101.67%之间。我们相信，我们的新型平台能够实现对可溶性癌症蛋白生物标志物的灵敏、多重和特异性检测，将极大地有益于癌症治疗并改善患者的预后。
