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

Journal: Frontiers in Microbiology

Article Title: A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration

doi: 10.3389/fmicb.2020.590736

Figure Lengend Snippet: PRBCHIP, an antibody microarray for detecting perchlorate reducing bacteria. (A) Schematic of the antibody printing pattern layout (by triplicate) in the PRBCHIP as indicated in and in Section “Materials and Methods.” Empty circles correspond to a serial dilution of a fluorescent antibody printed as a control for fluorescence (marked with a yellow dashed line); circles 1–20 represent antibodies raised against different strains; circles 21–24 indicate antibodies produced against proteins (red rectangles); P1–P20 indicate their corresponding pre-immune antibodies (marked with a continuous yellow line); circles labeled as B (BSA) and PP (Protein Printing Buffer) were used as negative control spots. (B,C) Images obtained after fluorescence sandwich microarray immunoassay (FSMI) with PRBCHIP by using cell lysates of Magnetospirillum bellicus VDY (B) and Dechlorobacter hydrogenophilus LT-1 (C) strains as sample. Immunoassays were revealed with anti- M. bellicus VDY (L2C1) and anti- D. hydrogenophilus LT- 1 (L4C1) antibodies, respectively. (D,E) Images obtained for the same samples and revealed with TOP-PRB fluorescent mix, made up of all anti-PRB antibodies shown in plus A-Cld and A-PCR antibodies. Red and white spots are fluorescence signals corresponding to positive immunodetections.

Article Snippet: The microarray images obtained with different immunogen dilutions were quantified and calibration curves were produced ( ) to determine the limit of detection (LOD) for each antibody ( ).

Techniques: Microarray, Bacteria, Serial Dilution, Control, Fluorescence, Produced, Labeling, Negative Control

Journal: Frontiers in Microbiology

Article Title: A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration

doi: 10.3389/fmicb.2020.590736

Figure Lengend Snippet: Testing the PRBCHIP sensitivity by fluorescent sandwich microarray immunoassay (FSMI). We assayed serial dilutions of cell cultures as samples for FSMI using their corresponding fluorescent antibodies or the TOP-PRB mix. The fluorescent signals in the microarray were quantified and plotted against the sample concentration. (A) Example of the calibration curve for antibody L1C1 raised against Azospira suillum PS and revealed with its tracer antibody and (B) with the TOP-PRB mix, respectively. When Azospira suillum PS was used as tested sample, besides L1C1 (black circles) and L1S2 (white circles), also L8C1 (triangles) showed a comparable signal at high sample concentrations. (C) Calibration curve for the antibody L4C1 raised against D. hydrogenophilus LT-1 revealed with its tracer antibody and (D) revealed with the TOP-PRB mix. In this case, only L4C1 showed positive immunodetections.

Article Snippet: The microarray images obtained with different immunogen dilutions were quantified and calibration curves were produced ( ) to determine the limit of detection (LOD) for each antibody ( ).

Techniques: Microarray, Concentration Assay

Journal: Frontiers in Microbiology

Article Title: A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration

doi: 10.3389/fmicb.2020.590736

Figure Lengend Snippet: Testing the specificity of the antibodies with the PRBCHIP. Immunogens corresponding to whole-cell extracts of each perchlorate reducing strain, its EPS fractions and proteins were tested by FSMI with its corresponding fluorescent antibody at its appropriate working dilution (see section “Materials and Methods”). (A) Specificities and cross-reactions between different antibodies and immunogens. Microarray images were scanned for fluorescence, quantified and plotted in a heatmap. Each column corresponds to a single PRBCHIP assay with the corresponding strain or protein tested (see section “Materials and Methods”) as the immunogen, while each row represents the antibody code for each printed antibody (as in ) on the microarray. (B) Antibody graph G of 16 nodes (antibodies with low performance were not considered for this analysis) and 28 links associated with our antibody microarray. Each node represents one antibody. The link (arrow) from antibody j to antibody i represents cross-reactivity of weight G ij , where G ij is the extent of cross-reactivity between antibodies i and j referred to the cognate immunogen of antibody j . Self-loops ( G jj = 1) are not shown for clarity.

Article Snippet: The microarray images obtained with different immunogen dilutions were quantified and calibration curves were produced ( ) to determine the limit of detection (LOD) for each antibody ( ).

Techniques: Microarray, Fluorescence

Journal: Frontiers in Microbiology

Article Title: A Multiplex Immunosensor for Detecting Perchlorate-Reducing Bacteria for Environmental Monitoring and Planetary Exploration

doi: 10.3389/fmicb.2020.590736

Figure Lengend Snippet: Deconvolution method applied to two complex natural samples by sandwich microarray immunoassay (FSMI). (A) Deconvolution of the American Pacific sample 140-10_07/07 and (B) deconvolution of the Lost Hammer sediment sample (LH-Sed) after performing both immunoassays with the mixture of all fluorescent-labeled tracer antibodies (TOP-PRB mix). Filled black bars represent the experimental fluorescence intensities F and red bars represent the deconvoluted signals F ′ [see (40–41) for details on obtaining F ′ from F and the matrix G associated to the antibody graph G]. Note that the experimental fluorescence intensities are ≥0 (as they were directly obtained from the FSMI), while the deconvoluted signals might be positive, zero or negative. Analyzing whether the experimental signal of each antibody is positive or close to zero and whether its deconvoluted signal is positive, zero or negative, we obtain a code for each organism (in bold) that yields reliable information about its existence or not in the sample [see supporting information of ].

Article Snippet: The microarray images obtained with different immunogen dilutions were quantified and calibration curves were produced ( ) to determine the limit of detection (LOD) for each antibody ( ).

Techniques: Microarray, Labeling, Fluorescence

Journal: Ecotoxicology (London, England)

Article Title: Application of physiologically based modelling and transcriptomics to probe the systems toxicology of aldicarb for Caenorhabditis elegans (Maupas 1900)

doi: 10.1007/s10646-010-0591-z

Figure Lengend Snippet: Scores plot for PC1 and PC2 from a PCA of normalised whole genome microarray data for C. elegans exposed to control or 16 mg/l aldicarb

Article Snippet: Acquired microarray images were quantified using ImaGene TM (Biodiscovery Inc., CA, USA) using the default flagging and segmentation settings.

Techniques: Microarray, Control

Journal:

Article Title: A protein multiplex microarray substrate with high sensitivity and specificity

doi: 10.1016/j.jim.2010.10.005

Figure Lengend Snippet: The Multiplex Microarray format is as follows: A1= Hepatitis B Core, A2 = Hepatitis C Core, A3 = HTLV-I, A4 = HTLV-II, B1 = T. Palladium, B2 = HIV-1 IIIB, B3 = HIV-2, B4 = Human Serum Albumin, 25 μg/μl packed colloid C1 = Human IgG, 25 μg/μl packed colloid, C2 = Human IgG, 12 μg/μl packed colloid, C3 = Human IgG, 6μg/μl packed colloid and C4 = Human Serum Albumin, 25 μg/μl packed colloid

Article Snippet: Presented in , are developed microarray images that were prepared by conventional direct spotting with the nitrocellulose colloid technology (A1, A2) and onto three commercial substrates: Prolinx (B), S&S FAST (C) and Telechem strepavadin (D).

Techniques: Multiplex Assay, Microarray

Journal:

Article Title: A protein multiplex microarray substrate with high sensitivity and specificity

doi: 10.1016/j.jim.2010.10.005

Figure Lengend Snippet: Light microscope (40X) images of developed colloidal microarrays. The multiplex microarray format is as follows: Each spot in triplicate, Row 1 = anti-norovirus, Row 2 = anti-adenovirus 41 (type specific), Row 3 = anti-adenovirus 40 (type specific), Row 4 = anti-hexon of adenovirus (group specific), Row 5 anti-astrovirus and Row 6 anti-rotavirus.

Article Snippet: Presented in , are developed microarray images that were prepared by conventional direct spotting with the nitrocellulose colloid technology (A1, A2) and onto three commercial substrates: Prolinx (B), S&S FAST (C) and Telechem strepavadin (D).

Techniques: Light Microscopy, Multiplex Assay, Microarray

Journal: Journal of Cancer

Article Title: Construction and validation of a comprehensive metabolism-associated prognostic model for predicting survival and immunotherapy benefits in ovarian cancer

doi: 10.7150/jca.100796

Figure Lengend Snippet: The comparison of levels of signature genes expression in tumor and normal controls. (A) Differences of expression levels of the four signature genes among the normal ovarian tissues and Ovarian cancer tissues (TCGA and GTEx database). (B) The relative mRNA levels of the four signature genes between normal ovarian cell lines and OV cell lines. (C) The prognostic value of a 4 sigurenaure gene for patients in TCGA has been confirmed through Kaplan-Meier analysis. (D) The immunohistochemical staining of tissue microarray shows 4 signature genes expressions on protein level between OV and normal control tissues.

Article Snippet: OV and normal ovarian tissue microarray (ZL-OVA961) were bought from ShangHai Zhuoli Biotech Company (China).

Techniques: Comparison, Expressing, Immunohistochemical staining, Staining, Microarray, Control