Department of Electro-Mechanical Systems Engineering, Korea University, Sejong, Chungnam 30019, Republic of Korea.
Department of Biomicrosystem Technology, Korea University, Seoul 136-701, Republic of Korea; Global Research Laboratory, Department of Biochemistry and Molecular Biology, Korea University College of Medicine, Seoul 136-713, Republic of Korea.
Biosens Bioelectron. 2017 Apr 15;90:238-244. doi: 10.1016/j.bios.2016.11.055. Epub 2016 Nov 25.
Development of CNT-based CD4 T cell imunosensors remains in its infancy due to the poor immobilization efficiency, lack of reproducibility, and difficulty in providing linear quantification. Here, we developed a fully-integrated single wall carbon nanotube (SWCNT)-based immunosensor capable of selective capture and linear quantification of CD4 T cells with greater dynamic range. By employing repeated two-step oxygen (O) plasma treatment processes with 35 days of recovery periods, we achieved the enhanced functionalization of the CNT surface and the removal of the byproduct of spray-coated SWCNTs that hinders charge transfer and stable CD4 T cell sensing. As a result, a linear electrochemical signal was generated in direct proportion to the bound cells. The slope of a SWCNT electrode in a target concentration range (10~10cells/mL) was 4.55×10μA per concentration decade, with the lowest detection limit of 1×10cells/mL. Since the reduced number of CD4 T cell counts in patients' peripheral blood corresponds to the progression of HIV disease, our CD4 T cell-immunosensor provides a simple and low-cost platform which can fulfill the requirement for the development of point-of-care (POC) diagnostic technologies for human immunodeficiency virus (HIV) patients in resource-limited countries.
由于较差的固定化效率、缺乏重现性以及难以进行线性定量,基于 CNT 的 CD4 T 细胞免疫传感器的发展仍处于起步阶段。在此,我们开发了一种完全集成的基于单壁碳纳米管 (SWCNT) 的免疫传感器,能够选择性捕获和线性定量 CD4 T 细胞,具有更大的动态范围。通过采用重复的两步氧(O)等离子体处理工艺,并在每个处理步骤之间留出 35 天的恢复期,我们实现了 CNT 表面的增强功能化,并去除了阻碍电荷转移和稳定 CD4 T 细胞检测的喷涂 SWCNTs 的副产物。结果,与结合细胞成正比的线性电化学信号得以产生。在目标浓度范围内(10~10cells/mL),SWCNT 电极的斜率为每浓度十年 4.55×10μA,最低检测限为 1×10cells/mL。由于患者外周血中 CD4 T 细胞数量的减少与 HIV 疾病的进展相对应,因此我们的 CD4 T 细胞免疫传感器提供了一个简单且低成本的平台,可满足资源有限国家开发针对人类免疫缺陷病毒 (HIV) 患者的即时护理 (POC) 诊断技术的需求。