Yun Kwang-Seok, Lee Dohoon, Kim Hak-Sung, Yoon Euisik
Department of Electronic Engineering, Sogang University, Seoul, Mapo-gu, Korea.
Methods Mol Biol. 2009;544:53-67. doi: 10.1007/978-1-59745-483-4_5.
This chapter introduces the demonstration of specific antibody detection by using a microbead-based assay with quantum dot (QD) fluorescence on a polydimethylsiloxane (PDMS) microfluidic chip. The microfluidic chip is designed to isolate a single microbead where the binding reaction of antibodies occurs on the surface. The microfluidic chip is fabricated on a glass substrate using a transparent silicone elastomer, PDMS, for easy access of monitoring and flexible gate operations to capture the single microbead. For antibody detection, a sequence of functionalized assays has been performed in the fabricated chip, including the capturing of microbeads, antibody injection into a microchamber, quantum dot injection, and fluorescence detection. Various concentrations of human IgG antibodies have been introduced to bind to a single microbead captured and isolated inside a designated microchamber in a small volume of 75 pL. Fluorescence detection is monitored using a CCD camera after the second binding with the QDs conjugated with anti-human IgG. In this experiment, a human IgG antibody concentration below 0.1 microg/mL has been successfully detected.
本章介绍了一种基于微珠的检测方法,该方法利用量子点(QD)荧光在聚二甲基硅氧烷(PDMS)微流控芯片上进行特定抗体检测的演示。微流控芯片的设计目的是分离单个微珠,抗体的结合反应在其表面发生。微流控芯片是在玻璃基板上使用透明硅橡胶PDMS制造的,便于进行监测和灵活的门操作以捕获单个微珠。对于抗体检测,在制造的芯片中进行了一系列功能化检测,包括微珠捕获、将抗体注入微腔、量子点注入和荧光检测。已引入不同浓度的人IgG抗体,使其与捕获并隔离在75 pL小体积指定微腔内的单个微珠结合。在与抗人IgG偶联的量子点进行第二次结合后,使用电荷耦合器件(CCD)相机监测荧光检测。在本实验中,已成功检测到低于0.1μg/mL的人IgG抗体浓度。