Department of Laboratory Medicine, The First Affiliated Hospital of Xiamen University, Xiamen, 361005, Fujian, China.
Department of Clinical Medicine, Fujian Medical University, Fuzhou, 350108, Fujian, China.
Anal Bioanal Chem. 2019 Oct;411(26):6837-6845. doi: 10.1007/s00216-019-02049-w. Epub 2019 Aug 30.
A new near-infrared-based photothermal immunosensing strategy was developed for the sensitive and feasible detection of human chorionic gonadotropin (HCG) by use of a Prussian blue nanoparticle-based photothermal conversion system. Prussian blue nanospheres synthesized by the one-pot method were used for the labeling of anti-HCG detection antibody. A sandwich-type immunoreaction was initially conducted on a monoclonal anti-HCG antibody-coated microplate with a nanoparticle-labeled signal antibody. Accompanying formation of the sandwiched immunocomplex, Prussian blue nanospheres caused photothermal conversion under 980-nm laser irradiation, thereby resulting in an increase of the temperature of the detection system measured by a portable digital thermometer. The properties and factors influencing the analytical performance of the photothermal immunoassay were studied in detail. Under the optimal conditions, the Prussian blue nanoparticle-based photothermal immunoassay exhibited good temperature responses relative to target HCG concentrations within the dynamic range of 0.01-100 ng mL at a low detection limit of 5.8 pg mL. This system also displayed good anti-interference behavior with regard to other cancer biomarkers, good reproducibility, and relatively long storage stability. The method accuracy was evaluated for analysis of human serum specimens, giving results that matched well with those obtained with a commercial HCG enzyme-linked immunosorbent assay kit. Importantly, this protocol is promising for advanced development of photothermal immunoassays. Graphical abstract.
一种新的基于近红外光的光热免疫传感策略被开发出来,用于通过普鲁士蓝纳米粒子光热转换系统灵敏且可行地检测人绒毛膜促性腺激素(HCG)。采用一锅法合成的普鲁士蓝纳米球被用于标记抗 HCG 检测抗体。在涂有单克隆抗 HCG 抗体的微孔板上,首先进行夹心型免疫反应,并用纳米球标记的信号抗体。伴随免疫复合物的形成,普鲁士蓝纳米球在 980nm 激光照射下引起光热转换,从而导致便携式数字温度计测量的检测系统温度升高。详细研究了光热免疫分析的性能和影响因素。在最佳条件下,基于普鲁士蓝纳米粒子的光热免疫分析在 5.8pg/mL 的低检测限下,对 0.01-100ng/mL 范围内的目标 HCG 浓度表现出良好的温度响应。该系统对其他癌症生物标志物也表现出良好的抗干扰行为、良好的重现性和相对较长的储存稳定性。该方法的准确性通过分析人血清标本进行了评估,结果与商业 HCG 酶联免疫吸附测定试剂盒的结果非常吻合。重要的是,该方案有望为光热免疫分析的进一步发展提供参考。