Membrane-binding peptides for extracellular vesicles on-chip analysis
Membrane-binding peptides for extracellular vesicles on-chip analysis
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DOI：
10.1080/20013078.2020.1751428
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发表时间：
2020-01-01
影响因子：
16
通讯作者：
Cretich, Marina
中科院分区：
文献类型：
--
作者：
Gori, Alessandro;Romanato, Alessandro;Cretich, Marina
小细胞外囊泡（sEV）在细胞外环境中呈现相当独特的脂质膜特征。这些包括高曲率，脂质包装缺陷和相对丰富的脂质，如磷脂酰丝氨酸和神经酰胺。因此，sEV膜可以被认为是一种“通用”标记物，替代或补充传统的、特征性的、表面相关的蛋白质。在这里，我们介绍了使用膜传感肽作为新的，高效的配体，直接集成sEV捕获和分析的微阵列平台。通过无标记、单颗粒计数和大小测定以及使用荧光抗CD 9/抗CD 63/抗CD 81抗体进行荧光共定位免疫染色来分析样品。肽表现为选择性但一般的sEV诱饵，并显示出比抗四跨膜蛋白抗体更高的结合能力。洞察表面化学的最佳肽性能进行了讨论，因为捕获效率是严格绑定到探针表面取向的影响。我们预计，这类新的配体，也由于合成肽的多功能性和有限的成本，可能会大大丰富EV分析的分子工具箱。
Small extracellular vesicles (sEVs) present fairly distinctive lipid membrane features in the extracellular environment. These include high curvature, lipid-packing defects and a relative abundance in lipids such as phosphatidylserine and ceramide. sEV membrane could be then considered as a "universal" marker, alternative or complementary to traditional, characteristic, surface-associated proteins. Here, we introduce the use of membrane-sensing peptides as new, highly efficient ligands to directly integrate sEV capturing and analysis on a microarray platform. Samples were analysed by label-free, single-particle counting and sizing, and by fluorescence co-localisation immune staining with fluorescent anti-CD9/anti-CD63/anti-CD81 antibodies. Peptides performed as selective yet general sEV baits and showed a binding capacity higher than anti-tetraspanins antibodies. Insights into surface chemistry for optimal peptide performances are also discussed, as capturing efficiency is strictly bound to probes surface orientation effects. We anticipate that this new class of ligands, also due to the versatility and limited costs of synthetic peptides, may greatly enrich the molecular toolbox for EV analysis.