Kim Hyung-Mo, Oh Chiwoo, An Jaehyun, Baek Seungki, Bock Sungje, Kim Jaehi, Jung Heung-Su, Song Hobeom, Kim Jung-Won, Jo Ahla, Kim Dong-Eun, Rho Won-Yeop, Jang Jin-Young, Cheon Gi Jeong, Im Hyung-Jun, Jun Bong-Hyun
Department of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Korea.
Department of Applied Bioengineering, Graduate School of Convergence Science and Technology, Seoul National University, Seoul 16229, Korea.
Nanomaterials (Basel). 2021 Mar 18;11(3):768. doi: 10.3390/nano11030768.
Exosomes are attracting attention as new biomarkers for monitoring the diagnosis and prognosis of certain diseases. Colorimetric-based lateral-flow assays have been previously used to detect exosomes, but these have the disadvantage of a high limit of detection. Here, we introduce a new technique to improve exosome detection. In our approach, highly bright multi-quantum dots embedded in silica-encapsulated nanoparticles (M-QD-SNs), which have uniform size and are brighter than single quantum dots, were applied to the lateral flow immunoassay method to sensitively detect exosomes. Anti-CD63 antibodies were introduced on the surface of the M-QD-SNs, and a lateral flow immunoassay with the M-QD-SNs was conducted to detect human foreskin fibroblast (HFF) exosomes. Exosome samples included a wide range of concentrations from 100 to 1000 exosomes/µL, and the detection limit of our newly designed system was 117.94 exosome/μL, which was 11 times lower than the previously reported limits. Additionally, exosomes were selectively detected relative to the negative controls, liposomes, and newborn calf serum, confirming that this method prevented non-specific binding. Thus, our study demonstrates that highly sensitive and quantitative exosome detection can be conducted quickly and accurately by using lateral immunochromatographic analysis with M-QD-SNs.
外泌体作为监测某些疾病诊断和预后的新型生物标志物正受到关注。基于比色法的侧向流动分析先前已用于检测外泌体,但这些方法具有检测限高的缺点。在此,我们介绍一种改进外泌体检测的新技术。在我们的方法中,将嵌入二氧化硅包封纳米颗粒(M-QD-SNs)中的高亮度多量子点应用于侧向流动免疫分析方法,以灵敏地检测外泌体,这些纳米颗粒尺寸均匀且比单量子点更亮。在M-QD-SNs表面引入抗CD63抗体,并进行M-QD-SNs的侧向流动免疫分析以检测人包皮成纤维细胞(HFF)外泌体。外泌体样品浓度范围为100至1000个外泌体/微升,我们新设计系统的检测限为117.94个外泌体/微升,比先前报道的检测限低11倍。此外,相对于阴性对照、脂质体和新生牛血清,外泌体被选择性地检测到,证实该方法可防止非特异性结合。因此,我们的研究表明,通过使用M-QD-SNs的侧向免疫色谱分析,可以快速准确地进行高灵敏度和定量的外泌体检测。