Department of Physics Education, Chosun University, Gwangju, 61452, Republic of Korea.
Department of Biomedical Science, Chosun University, Gwangju, 61452, Republic of Korea.
Anal Biochem. 2023 Jul 15;673:115161. doi: 10.1016/j.ab.2023.115161. Epub 2023 May 16.
Exosomes are potential biomarkers for disease diagnosis and treatment, as well as drug carriers. However, as their isolation and detection remain critical issues, convenient, rapid, low-cost, and effective methods are necessary. In this study, we present a rapid and simple method for directly capturing and analyzing exosomes from complex cell culture media using CaTiO:Eu@FeO multifunctional nanocomposites. The CaTiO:Eu@FeO nanocomposites were prepared by high-energy ball-milling and used to isolate exosomes by binding CaTiO:Eu@FeO nanocomposites and the hydrophilic phosphate head of the exosome phospholipids. Notably, the developed CaTiO:Eu@FeO multifunctional nanocomposites achieved results comparable with those of commercially available TiO and were separated using a magnet within 10 min. Moreover, we report a surface-enhanced Raman scattering (SERS)-based immunoassay for detecting the exosome biomarker CD81. Gold nanorods (Au NRs) were modified with detection antibodies, and antibody-conjugated Au NRs were labeled with 3, 3, diethylthiatricarbocyanine iodide (DTTC) as the SERS tags. A method combining magnetic separation and SERS was developed to detect exosomal biomarker CD81. The results of this study demonstrate the feasibility of this new technique as a useful tool for exosome isolation and detection.
外泌体是疾病诊断和治疗以及药物载体的潜在生物标志物。然而,由于其分离和检测仍然是关键问题,因此需要方便、快速、低成本和有效的方法。在本研究中,我们提出了一种使用 CaTiO:Eu@FeO 多功能纳米复合材料直接从复杂细胞培养介质中捕获和分析外泌体的快速简便方法。CaTiO:Eu@FeO 纳米复合材料通过高能球磨制备,并通过结合 CaTiO:Eu@FeO 纳米复合材料和亲水磷酸头的外泌体磷脂来分离外泌体。值得注意的是,开发的 CaTiO:Eu@FeO 多功能纳米复合材料与市售 TiO 的结果相当,并在 10 分钟内通过磁铁分离。此外,我们报告了一种基于表面增强拉曼散射(SERS)的免疫测定法,用于检测外泌体生物标志物 CD81。金纳米棒(Au NRs)用检测抗体修饰,并用 3,3,二乙基噻二碳菁碘化物(DTTC)作为 SERS 标记物标记抗体偶联的 Au NRs。开发了一种结合磁分离和 SERS 的方法来检测外泌体生物标志物 CD81。该研究结果证明了这项新技术作为外泌体分离和检测有用工具的可行性。