Program in Nanoscience and Technology, Graduate School of Convergence Science and Technology , Seoul National University , Seoul 08826 , Republic of Korea.
Biomacromolecules. 2019 Sep 9;20(9):3392-3398. doi: 10.1021/acs.biomac.9b00641. Epub 2019 Aug 6.
Exosomes are extracellular vesicles (EVs) that have attracted attention because of their important biological roles in intercellular communication and transportation of various biomolecules, including proteins and genetic materials. However, due to difficulties in their selective capture and detection, further application of exosomes remains challenging. To detect EVs, we fabricated a liposomal biosensor based on polydiacetylene (PDA), a conjugate polymer that has been widely used in sensing applications derived from its unique optical properties. To confer selectivity and sensitivity to the sensory material, antibodies targeting CD63, a membrane protein exclusively found in exosomes, were attached to the PDA liposomes and phospholipid molecules were incorporated into the PDA vesicles. Signal analysis derived from PDA liposomes for exosome detection and quantification was performed by observing colorimetric changes triggered by the ligand-receptor interaction of PDA vesicles. Visual, UV-visible, and fluorescence spectroscopic methods were used to obtain signals from the PDA lipid immunosensor, which achieved a detection limit of 3 × 10 vesicles/mL, the minimum concentration that can be used in practical applications. The strategies used in the system have the potential to expand into the field of dealing with exosomes.
外泌体是细胞外囊泡 (EVs),由于其在细胞间通讯和运输各种生物分子(包括蛋白质和遗传物质)方面的重要生物学作用而受到关注。然而,由于其选择性捕获和检测的困难,外泌体的进一步应用仍然具有挑战性。为了检测 EVs,我们基于聚二乙炔 (PDA) 构建了脂质体生物传感器,聚二乙炔是一种共轭聚合物,由于其独特的光学特性,已广泛应用于传感应用中。为了赋予感测材料选择性和敏感性,针对 CD63(一种仅在外泌体中发现的膜蛋白)的抗体被连接到 PDA 脂质体上,并将磷脂分子掺入到 PDA 囊泡中。通过观察 PDA 囊泡的配体-受体相互作用引发的比色变化来对用于外泌体检测和定量的 PDA 脂质体进行信号分析。使用可见、紫外-可见和荧光光谱法从 PDA 脂质体免疫传感器中获取信号,该传感器的检测限为 3×10 个囊泡/mL,这是实际应用中可以使用的最小浓度。该系统中使用的策略有可能扩展到外泌体处理领域。