[15a-WL1_201-1~11] 12.6 Nanobiotechnology

11:45 AM - 12:00 PM JST(2:45 AM - 3:00 AM UTC)

[15a-WL1_201-11] Membrane Protein Analysis Using a Plasmonic Chip of Single Extracellular Vesicles with Lipid Membrane Staining

Tatsuki Iwamura¹, 〇Yasunori Nawa¹, Rina Nagayama¹, Hajime Shigeto², Shohei Yamamura², Keiko Tawa¹ (1.Kwansei Gakuin Univ., 2.AIST)

Keywords:

Exosome,Enhanced fluorescence,Plasmonic chip

Exosomes, a class of extracellular vesicles (EVs) secreted by cells, play important roles in intercellular communication. However, their sub-micrometer size and weak optical signals make single-EV analysis by conventional optical microscopy challenging. To address this limitation, we improved detection sensitivity using a plasmonic chip–based fluorescence enhancement technique. In this study, we further adopted lipid membrane staining instead of a sandwich immunoassay to improve the quantitative accuracy of single-EV analysis. A plasmonic chip functionalized with capture antibodies targeting CD63, CD326, and CD151 as EV surface markers enabled the detection of lipid-labeled single EVs as fluorescent spots. In observations of EVs secreted from MCF-7 breast cancer cells, CD151, which is not expressed in normal cells, was clearly detected, and its secretion level exhibited culture duration dependence similar to that of CD63 and CD326.