Department of Chemistry, Seoul National University, Seoul 08826, Republic of Korea.
InSol Co., Ltd., Yangjae-daero 85-gil, Gangdong-gu, Seoul 05408, Republic of Korea.
Biosens Bioelectron. 2018 Apr 15;102:121-128. doi: 10.1016/j.bios.2017.11.027. Epub 2017 Nov 7.
Bead-based multiplex immunoassays for common use require enhanced sensitivity and effective prevention of non-specific adsorption, as well as miniaturization of the detection device. In this work, we have implemented virus-tethered gold microspheres for multiplex immunoassay applications, employing a DC impedance-based flow cytometer as a detection element. The advantages of virus-tethered gold microspheres, including excellent prevention of non-specific adsorption, are extended to signal enhancement arising from the large quantity of antibody loading on each virion, and to flexible movement of filamentous virus. Individual virus-tethered beads generate their own DC impedance and fluorescence signals, which are simultaneously detected by a chip-based microfluidic flow cytometer. This system successfully realized multiplex immunoassays involving four biomarkers: cardiac troponin I (cTnI), prostate specific antigen (PSA), creatine kinase MB (CK-MB), and myoglobin in undiluted human sera, elevating sensitivity by up to 5.7-fold compared to the beads without virus. Constructive integration between filamentous virus-tethered Au-layered microspheres and use of a microfluidic cytometer suggests a promising strategy for competitive multiplex immunoassay development based on suspension arrays.
基于珠体的多重免疫分析通常需要提高灵敏度和有效防止非特异性吸附，以及检测装置的小型化。在这项工作中，我们已经实现了用于多重免疫分析应用的病毒连接金微球，采用基于直流阻抗的流动细胞仪作为检测元件。病毒连接金微球的优点，包括优异的防止非特异性吸附，扩展到信号增强由于每个病毒粒子上加载的大量抗体，以及丝状病毒的灵活运动。单个病毒连接珠体产生自己的直流阻抗和荧光信号，这两种信号都由基于芯片的微流控流动细胞仪同时检测。该系统成功地实现了涉及四种生物标志物的多重免疫分析：心肌肌钙蛋白 I（cTnI）、前列腺特异性抗原（PSA）、肌酸激酶 MB（CK-MB）和肌红蛋白在未稀释的人血清中，与没有病毒的珠体相比，灵敏度提高了 5.7 倍。丝状病毒连接 Au 层微球的建设性集成和微流控细胞仪的使用表明，基于悬浮阵列的竞争多重免疫分析开发具有有前途的策略。
