TY - JOUR
T1 - Membrane-binding peptides for extracellular vesicles on-chip analysis
AU - Gori, Alessandro
AU - Romanato, Alessandro
AU - Greta, Bergamaschi
AU - Strada, Alessandro
AU - Gagni, Paola
AU - Frigerio, Roberto
AU - Brambilla, Dario
AU - Vago, Riccardo
AU - Galbiati, Silvia
AU - Picciolini, Silvia
AU - Bedoni, Marzia
AU - Daaboul, George G.
AU - Chiari, Marcella
AU - Cretich, Marina
PY - 2020/4/1
Y1 - 2020/4/1
N2 - 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.
AB - 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.
KW - Extracellular vesicles
KW - membrane binding
KW - membrane curvature
KW - microarrays
KW - peptides
UR - http://www.scopus.com/inward/record.url?scp=85083566347&partnerID=8YFLogxK
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U2 - 10.1080/20013078.2020.1751428
DO - 10.1080/20013078.2020.1751428
M3 - Article
AN - SCOPUS:85083566347
VL - 9
JO - Journal of Extracellular Vesicles
JF - Journal of Extracellular Vesicles
SN - 2001-3078
IS - 1
M1 - 1751428
ER -