Li Chao, Yang Yucai, Wu Dan, Li Tianqi, Yin Yongmei, Li Genxi
State Key Laboratory of Pharmaceutical Biotechnology , Collaborative Innovation Center of Chemistry for Life Sciences , Department of Biochemistry , Nanjing University , Nanjing 210093 , China . Email:
Department of Oncology , The First Affiliated Hospital of Nanjing Medical University , Nanjing 210029 , China.
Chem Sci. 2016 May 1;7(5):3011-3016. doi: 10.1039/c5sc04256a. Epub 2016 Feb 3.
An enzyme-linked immunosorbent assay that is dependent on enzyme amplification has dominated the current field of protein detection; however, limited multiple detection ability and susceptible enzymatic reactions, and low sensitivity may severely hinder its application. Here, we report a new signal amplification scheme based on allochroic molecule modified carboxyl graphene oxide (cGO), which can be used to develop a multicolor immunoassay named as allochroic-cGO linked immunosorbent assay (ALISA). Thanks to high adsorption levels and a wide selection of allochroic molecules, the simultaneous colorimetric detection of diagnostic biomarkers at a picogram level can be successfully achieved for the first time. In addition, the color change triggered by acidic or basic water can provide a simple, rapid, stable and economical signal output, further meeting the growing biodetection requirements. Moreover, with the help of ALISA, we demonstrate that the combined detection of three tumor biomarkers, including carcino-embryonic antigen, neuron-specific enolase, and cytokeratin-19 fragment, is more valuable for differentiating lung cancer patients than the detection of a single biomarker, further manifesting the superiority of ALISA. All in all, this straightforward approach not only opens up new prospects for multicolor immunoassays, but also has great potential for applications in resource-constrained settings.
一种依赖酶扩增的酶联免疫吸附测定法主导了当前的蛋白质检测领域；然而，其有限的多重检测能力、易受影响的酶促反应以及低灵敏度可能会严重阻碍其应用。在此，我们报告了一种基于变色分子修饰的羧基氧化石墨烯（cGO）的新信号放大方案，该方案可用于开发一种名为变色 - cGO 联免疫吸附测定法（ALISA）的多色免疫测定法。由于高吸附水平和多种变色分子可供选择，首次成功实现了皮克级诊断生物标志物的比色法同时检测。此外，由酸性或碱性水引发的颜色变化可提供简单、快速、稳定且经济的信号输出，进一步满足不断增长的生物检测需求。此外，借助 ALISA，我们证明联合检测三种肿瘤生物标志物，包括癌胚抗原、神经元特异性烯醇化酶和细胞角蛋白 19 片段，比检测单一生物标志物在区分肺癌患者方面更有价值，进一步彰显了 ALISA 的优越性。总而言之，这种直接的方法不仅为多色免疫测定开辟了新前景，而且在资源有限的环境中具有巨大的应用潜力。
