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single-ev microarray imaging  (NanoView Biosciences)
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NanoView Biosciences single-ev microarray imaging
Characterization of RSV-Specific Peptide Engineered DC Exosomes. ( A ) Schematic illustration of ExoRelease beads prepared, peptide-engineered exosomes via MHC binding peptide capture, surface MHC peptide coating, and photo-release for harvesting. ( B ) SEM images showing the ExoRelease bead surface morphology during the exosome capture, surface engineering, and release of intact captured exosomes. The entire surface was covered by round-shape exosomes after capture which is significantly different than the bare surface after release of intact captured exosomes. ( C ) TEM images showing the morphology of harvested exosomes using ExoRelease beads approach, which is in uniform size and round shape. However, ultracentrifugation isolated vesicles are more heterogeneous with substantial particle aggregates. ( D ) The nanoparticle tracking analysis of harvested, peptide-engineered exosomes using ExoRelease bead approach, compared with ultracentrifugation isolated vesicles without peptide engineering. Here two peptides M and NS were prepared for exosome surface engineering. The peptide engineered exosomes are in much narrower size distribution. ( E ) Zeta potential analysis of surface engineered exosomes compared with ultracentrifugation isolated vesicles as the control group. The zeta potential from peptide engineered exosomes were slightly changed due to the surface engineering, but still maintain the good integrity. ( F ) NanoView <t>microarray</t> was used to determine the level of MHC-I the exosomes. Exosomes from LPS stimulated JAWS expressed more MHC-I on their surface. ( G ) The stacked bar chart showing the percentage distribution of total exosomes, total MHC-I on exosome surface, and total RSV peptides (M and NS) bound to exosome MHC-I, which is analyzed by bead-based flow cytometry. ( H ) The rate of engineered RSV peptides (M and NS) on exosome surfaces (n = 3, RSD < 5%).
Single Ev Microarray Imaging, supplied by NanoView Biosciences, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Characterization of RSV-Specific Peptide Engineered DC Exosomes. ( A ) Schematic illustration of ExoRelease beads prepared, peptide-engineered exosomes via MHC binding peptide capture, surface MHC peptide coating, and photo-release for harvesting. ( B ) SEM images showing the ExoRelease bead surface morphology during the exosome capture, surface engineering, and release of intact captured exosomes. The entire surface was covered by round-shape exosomes after capture which is significantly different than the bare surface after release of intact captured exosomes. ( C ) TEM images showing the morphology of harvested exosomes using ExoRelease beads approach, which is in uniform size and round shape. However, ultracentrifugation isolated vesicles are more heterogeneous with substantial particle aggregates. ( D ) The nanoparticle tracking analysis of harvested, peptide-engineered exosomes using ExoRelease bead approach, compared with ultracentrifugation isolated vesicles without peptide engineering. Here two peptides M and NS were prepared for exosome surface engineering. The peptide engineered exosomes are in much narrower size distribution. ( E ) Zeta potential analysis of surface engineered exosomes compared with ultracentrifugation isolated vesicles as the control group. The zeta potential from peptide engineered exosomes were slightly changed due to the surface engineering, but still maintain the good integrity. ( F ) NanoView microarray was used to determine the level of MHC-I the exosomes. Exosomes from LPS stimulated JAWS expressed more MHC-I on their surface. ( G ) The stacked bar chart showing the percentage distribution of total exosomes, total MHC-I on exosome surface, and total RSV peptides (M and NS) bound to exosome MHC-I, which is analyzed by bead-based flow cytometry. ( H ) The rate of engineered RSV peptides (M and NS) on exosome surfaces (n = 3, RSD < 5%).

Journal: Scientific Reports

Article Title: Development of surface engineered antigenic exosomes as vaccines for respiratory syncytial virus

doi: 10.1038/s41598-021-00765-x

Figure Lengend Snippet: Characterization of RSV-Specific Peptide Engineered DC Exosomes. ( A ) Schematic illustration of ExoRelease beads prepared, peptide-engineered exosomes via MHC binding peptide capture, surface MHC peptide coating, and photo-release for harvesting. ( B ) SEM images showing the ExoRelease bead surface morphology during the exosome capture, surface engineering, and release of intact captured exosomes. The entire surface was covered by round-shape exosomes after capture which is significantly different than the bare surface after release of intact captured exosomes. ( C ) TEM images showing the morphology of harvested exosomes using ExoRelease beads approach, which is in uniform size and round shape. However, ultracentrifugation isolated vesicles are more heterogeneous with substantial particle aggregates. ( D ) The nanoparticle tracking analysis of harvested, peptide-engineered exosomes using ExoRelease bead approach, compared with ultracentrifugation isolated vesicles without peptide engineering. Here two peptides M and NS were prepared for exosome surface engineering. The peptide engineered exosomes are in much narrower size distribution. ( E ) Zeta potential analysis of surface engineered exosomes compared with ultracentrifugation isolated vesicles as the control group. The zeta potential from peptide engineered exosomes were slightly changed due to the surface engineering, but still maintain the good integrity. ( F ) NanoView microarray was used to determine the level of MHC-I the exosomes. Exosomes from LPS stimulated JAWS expressed more MHC-I on their surface. ( G ) The stacked bar chart showing the percentage distribution of total exosomes, total MHC-I on exosome surface, and total RSV peptides (M and NS) bound to exosome MHC-I, which is analyzed by bead-based flow cytometry. ( H ) The rate of engineered RSV peptides (M and NS) on exosome surfaces (n = 3, RSD < 5%).

Article Snippet: We also used the single-EV microarray imaging technology from NanoView to directly determine the MHC-I expression level from prepared exosomes as well as their functional markers shown in Fig. F. The specific antibody capture on each NanoView chip spot allows the affinity capture of exosomes based on their surface markers for further multiplexed affinity probing.

Techniques: Binding Assay, Isolation, Zeta Potential Analyzer, Control, Microarray, Flow Cytometry