State Key Lab of Metal Matrix Composites, School of Materials Science and Engineering, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, PR China.
Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai Jiao Tong University, 1665 Kongjiang Road, Shanghai, 200092, PR China.
Biosens Bioelectron. 2020 Feb 15;150:111912. doi: 10.1016/j.bios.2019.111912. Epub 2019 Nov 21.
Aggregation-induced emission luminogens (AIEgens) have attracted considerable interest for application towards the development of various biosensors due to their unique optical properties. However, the major challenge associated with generating a suitable fluorescent signal for constructing an AIEgens-based immunoassay platform, is the complex surface modification and additional chemical reaction required to activate the AIE process. This work reports a novel AIEgens nanobeads-based fluorescence-linked immunosorbent assay (FLISA) platform wherein the fluorescent labels are hexaphenylsilole (HPS) nanobeads, which were synthesized through Shirasu porous glass (SPG) membrane emulsification method and could provide a strong, direct fluorescent signal without any pretreatment. Moreover, the particle-based signal amplification effect affords this platform significantly improved detection sensitivity for carcinoembryonic antigen (CEA) quantitation. Compared to FLISA which uses R-phycoerythrin (PE) or commercial green QDs nanobeads as fluorescent labels, this AIEgens nanobeads-based FLISA platform exhibits detection sensitivity improved up to 45-fold and 12-fold, respectively. Clinical validation experiments applying this AIEgens nanobeads-based FLISA immunoassay platform to analyze human serum samples produce results consistent with those obtained by the clinical gold-standard method, electrochemiluminescence immunoassay (ECLIA). The strong photobleaching resistance and excellent fluorescent stability of the HPS nanobeads negate the need for light shielding, which improves the efficiency and makes the operating conditions more comfortable. Thus, this AIEgens nanobeads-based FLISA platform, with attractive features including direct fluorescent signal generation and significant signal amplification, creates a new, versatile route for the application of AIEgens in biosensors and clinical diagnosis.
聚集诱导发光材料（AIEgens）由于其独特的光学性质，在开发各种生物传感器方面引起了相当大的兴趣。然而，对于构建基于 AIEgens 的免疫分析平台，产生合适的荧光信号的主要挑战是复杂的表面修饰和额外的化学反应，这些反应需要激活 AIE 过程。本工作报道了一种新型的基于 AIEgens 纳米珠的荧光免疫吸附测定（FLISA）平台，其中荧光标记物是六苯基硅咯（HPS）纳米珠，通过 Shirasu 多孔玻璃（SPG）膜乳化法合成，可提供无需任何预处理的强、直接荧光信号。此外，基于颗粒的信号放大效应，使该平台对癌胚抗原（CEA）定量的检测灵敏度显著提高。与使用 R-藻红蛋白（PE）或商业绿色 QDs 纳米珠作为荧光标记物的 FLISA 相比，这种基于 AIEgens 纳米珠的 FLISA 平台的检测灵敏度分别提高了 45 倍和 12 倍。应用这种基于 AIEgens 纳米珠的 FLISA 免疫分析平台分析人血清样本的临床验证实验结果与临床金标准方法（电化学发光免疫分析（ECLIA））一致。HPS 纳米珠的强光漂白抗性和优异的荧光稳定性消除了对遮光的需求，从而提高了效率并使操作条件更加舒适。因此，这种基于 AIEgens 纳米珠的 FLISA 平台具有直接荧光信号产生和显著信号放大的特点，为 AIEgens 在生物传感器和临床诊断中的应用开辟了一条新的、多功能的途径。
