Isolation:Article Title: Mucosal vaccination against SARS-CoV-2 using recombinant influenza viruses delivering self-assembling nanoparticles.
Article Snippet: Cells from lungs were treated with PharmLyse (BD) to lyse red blood cells prior to downstream staining. .. Single cells were isolated and stained with Aqua viability dye (Thermofisher) and Fc-blocked with anti-CD16/32 antibody (clone 93, Biolegend). .. Cells were then labelled with PE conjugated RBD probes, and the following antibodies: B220 BUV737 (clone RA3-6B2; BD), CD45 Cy7-APC (clone 30-F11; BD), IgD BUV 395 (clone 11-26c.2a; BD), GL7 AF488 (clone GL7; BioLegend), CD38 Cy7-PE (clone 90; BioLegend), Streptavidin BV786 (BD), CD3 BV786 (clone145-2C11; BioLegend), F4/ 80 BV786 (BM8; BioLegend).
Article Title: Mucosal vaccination against SARS-CoV-2 using recombinant influenza viruses delivering self-assembling nanoparticles.
Article Snippet: Flow cytometry data were processed using FlowJo v10. .. Single cells were isolated and stained with Aqua viability dye (Thermofisher) and Fc-blocked with anti-CD16/32 antibody (comp/ clone). .. Cells were then labelled with RBD-PE probes, and the following antibodies:, CD138 BB515 (clone 281–2, BD), CD98 BUV737 (clone H202–141, BD), B220 BV650 (clone RA3-6B2, BioLegend), CD45 Cy7APC (clone 30-F11; BD), IgD BUV395 (clone 11-26c.2a; BD), GL7 PacBlue (clone GL7, BioLegend), CD38 Cy7-PE (clone 90; BioLegend), Streptavidin BV510 (BD), CD3 BV510 (clone 17A2, BioLegend) F4/80 BV510 (clone T45–2342, BD).
Article Title: Modulation of germinal center and antibody dynamics via ipsilateral versus contralateral immunization against SARS-CoV-2.
Article Snippet: Flow cytometry data were processed using FlowJo v10 (TreeStar). .. Single-cell sorting and B cell receptor sequencing Isolated lymphocytes were stained with Aqua viability dye (Thermo Fisher Scientific) and Fc-blocked with anti-CD16/32 antibody (clone 93; BioLegend). .. Cells were then labeled with PE- and APC-conjugated S probes and the following antibodies: F4/80 BV786 (BM8; BioLegend; 1:150), CD3 BV786 (145- 2C11; BioLegend; 1:750), SA BV786 (BD), CD45 APC-Cy7 (30-F11; BD; 1:300), CD38 PeCy7 (90; BioLegend; 1:750), GL7 AF488 (GL7; BioLegend; 1:300), IgD PerCPCy5.5 (11- 26c2a; BD; 1:300), IgM BV421 (II/41; BD; 1:300), and B220 BV650 (RA3-6B2; BD; 1:300).
Staining:Article Title: Mucosal vaccination against SARS-CoV-2 using recombinant influenza viruses delivering self-assembling nanoparticles.
Article Snippet: Cells from lungs were treated with PharmLyse (BD) to lyse red blood cells prior to downstream staining. .. Single cells were isolated and stained with Aqua viability dye (Thermofisher) and Fc-blocked with anti-CD16/32 antibody (clone 93, Biolegend). .. Cells were then labelled with PE conjugated RBD probes, and the following antibodies: B220 BUV737 (clone RA3-6B2; BD), CD45 Cy7-APC (clone 30-F11; BD), IgD BUV 395 (clone 11-26c.2a; BD), GL7 AF488 (clone GL7; BioLegend), CD38 Cy7-PE (clone 90; BioLegend), Streptavidin BV786 (BD), CD3 BV786 (clone145-2C11; BioLegend), F4/ 80 BV786 (BM8; BioLegend).
Article Title: Mucosal vaccination against SARS-CoV-2 using recombinant influenza viruses delivering self-assembling nanoparticles.
Article Snippet: Flow cytometry data were processed using FlowJo v10. .. Single cells were isolated and stained with Aqua viability dye (Thermofisher) and Fc-blocked with anti-CD16/32 antibody (comp/ clone). .. Cells were then labelled with RBD-PE probes, and the following antibodies:, CD138 BB515 (clone 281–2, BD), CD98 BUV737 (clone H202–141, BD), B220 BV650 (clone RA3-6B2, BioLegend), CD45 Cy7APC (clone 30-F11; BD), IgD BUV395 (clone 11-26c.2a; BD), GL7 PacBlue (clone GL7, BioLegend), CD38 Cy7-PE (clone 90; BioLegend), Streptavidin BV510 (BD), CD3 BV510 (clone 17A2, BioLegend) F4/80 BV510 (clone T45–2342, BD).
Article Title: Deconvolution of Cargo Delivery and Immunogenicity Following Intranasal Delivery of mRNA Lipid Nanoparticle Vaccines
Article Snippet: Red blood cells from the lungs were lysed with PharmLyse (BD). .. Single cells were stained with Aqua viability dye (Thermo Fisher Scientific) and Fc-blocked with anti-CD16/CD32 antibodies. .. The cells were then labeled with OVA-PE and OVAAPC probes, and the following antibody panel: F4/80 BV786 (BM8; BioLegend), B220 BUV737 (RA3-6B2; BD), CD45 Cy7-APC (30-F11; BD), IgD BUV 395 (11-26c.2a; BD), GL7 AF488 (GL7; BioLegend), and CD38 Cy7-PE (clone 90; BioLegend).
Article Title: A novel chimeric coronavirus spike vaccine combining SARS-CoV-2 RBD and scaffold domains from HKU-1 elicits potent neutralising antibody responses
Article Snippet: .. Cells were stained with Aqua viability dye (Thermo Fisher) and Fc-blocked with anti-CD16/32 antibody (clone 93, BioLegend). .. Cells were then labelled with PE- and APC-conjugated spike probes, and the following antibodies at 4°C: F4/80 BV786 (BM8; BioLegend; 1:150), CD3 BV786 (145-2C11; BioLegend; 1:750), strepavidin BV786 (BD), CD45 APC-Cy7 (30-F11; BD; 1:300), CD38 PeCy7 (90; BioLegend; 1:750), GL7 AF488 (GL7; BioLegend; 1:300), IgD BUV395 (11-26c.2a; BD; 1:300), and B220 BUV737 (RA3-6B2; BD; 1:300).
Article Title: Potent neutralising monoclonal antibodies targeting the spike of NL63 coronavirus
Article Snippet: Bound antibody was detected using 1:20,000 dilution of HRP-conjugated anti-human IgG (Sigma) and plates developed using TMB substrate (Sigma), stopped using sulphuric acid and read at 450 nm. .. PBMCs were thawed and stained with Aqua viability dye (Thermo Fisher Scientific) and then surface stained with Spike probes, CD19 ECD (J3-119) (Beckman Coulter), IgD PE-Cy7 (IA6-2), IgG BV786 (G18-145), streptavidin BV510 (BD Biosciences), CD14 BV510 (M5E2), CD3 BV510 (OKT3), CD8a BV510 (RPA-T8), CD16 BV510 (3G8), and CD10 BV510 (HI10a) (BioLegend). .. Cells were washed twice with PBS containing 1% FCS and fixed with 1% formaldehyde (Polysciences), acquired on a BDLSR Fortessa using BD FACS Diva and analysed using FlowJo 10.
Article Title: Modulation of germinal center and antibody dynamics via ipsilateral versus contralateral immunization against SARS-CoV-2.
Article Snippet: .. Cells were stained with Aqua viability dye (Thermo Fisher) and Fcblocked with anti-CD16/32 antibody (clone 93; BioLegend). .. Cells were then labeled with PE- and APC-conjugated S probes and the following antibodies: F4/80 BV786 (BM8; BioLegend; 1:150), CD3 BV786 (145-2C11; BioLegend; 1:750), strepavidin BV786 (BD), CD45 APC-Cy7 (30-F11; BD; 1:300), CD38 PeCy7 (90; BioLegend; 1:750), GL7 AF4788 (GL7; BioLegend; 1:300), IgD BUV395 (11-26c.2a; BD; 1:300), B220 BUV737 (RA3-6B2; BD; 1:300), CD98 BV711 (H202-141; BD; 1:100), and CD138 BV421 (281-2; BD; 1:100).
Article Title: Modulation of germinal center and antibody dynamics via ipsilateral versus contralateral immunization against SARS-CoV-2.
Article Snippet: Flow cytometry data were processed using FlowJo v10 (TreeStar). .. Single-cell sorting and B cell receptor sequencing Isolated lymphocytes were stained with Aqua viability dye (Thermo Fisher Scientific) and Fc-blocked with anti-CD16/32 antibody (clone 93; BioLegend). .. Cells were then labeled with PE- and APC-conjugated S probes and the following antibodies: F4/80 BV786 (BM8; BioLegend; 1:150), CD3 BV786 (145- 2C11; BioLegend; 1:750), SA BV786 (BD), CD45 APC-Cy7 (30-F11; BD; 1:300), CD38 PeCy7 (90; BioLegend; 1:750), GL7 AF488 (GL7; BioLegend; 1:300), IgD PerCPCy5.5 (11- 26c2a; BD; 1:300), IgM BV421 (II/41; BD; 1:300), and B220 BV650 (RA3-6B2; BD; 1:300).
Recombinase Polymerase Amplification:Article Title: Potent neutralising monoclonal antibodies targeting the spike of NL63 coronavirus
Article Snippet: Bound antibody was detected using 1:20,000 dilution of HRP-conjugated anti-human IgG (Sigma) and plates developed using TMB substrate (Sigma), stopped using sulphuric acid and read at 450 nm. .. PBMCs were thawed and stained with Aqua viability dye (Thermo Fisher Scientific) and then surface stained with Spike probes, CD19 ECD (J3-119) (Beckman Coulter), IgD PE-Cy7 (IA6-2), IgG BV786 (G18-145), streptavidin BV510 (BD Biosciences), CD14 BV510 (M5E2), CD3 BV510 (OKT3), CD8a BV510 (RPA-T8), CD16 BV510 (3G8), and CD10 BV510 (HI10a) (BioLegend). .. Cells were washed twice with PBS containing 1% FCS and fixed with 1% formaldehyde (Polysciences), acquired on a BDLSR Fortessa using BD FACS Diva and analysed using FlowJo 10.
Single Cell:Article Title: Modulation of germinal center and antibody dynamics via ipsilateral versus contralateral immunization against SARS-CoV-2.
Article Snippet: Flow cytometry data were processed using FlowJo v10 (TreeStar). .. Single-cell sorting and B cell receptor sequencing Isolated lymphocytes were stained with Aqua viability dye (Thermo Fisher Scientific) and Fc-blocked with anti-CD16/32 antibody (clone 93; BioLegend). .. Cells were then labeled with PE- and APC-conjugated S probes and the following antibodies: F4/80 BV786 (BM8; BioLegend; 1:150), CD3 BV786 (145- 2C11; BioLegend; 1:750), SA BV786 (BD), CD45 APC-Cy7 (30-F11; BD; 1:300), CD38 PeCy7 (90; BioLegend; 1:750), GL7 AF488 (GL7; BioLegend; 1:300), IgD PerCPCy5.5 (11- 26c2a; BD; 1:300), IgM BV421 (II/41; BD; 1:300), and B220 BV650 (RA3-6B2; BD; 1:300).
Sequencing:Article Title: Modulation of germinal center and antibody dynamics via ipsilateral versus contralateral immunization against SARS-CoV-2.
Article Snippet: Flow cytometry data were processed using FlowJo v10 (TreeStar). .. Single-cell sorting and B cell receptor sequencing Isolated lymphocytes were stained with Aqua viability dye (Thermo Fisher Scientific) and Fc-blocked with anti-CD16/32 antibody (clone 93; BioLegend). .. Cells were then labeled with PE- and APC-conjugated S probes and the following antibodies: F4/80 BV786 (BM8; BioLegend; 1:150), CD3 BV786 (145- 2C11; BioLegend; 1:750), SA BV786 (BD), CD45 APC-Cy7 (30-F11; BD; 1:300), CD38 PeCy7 (90; BioLegend; 1:750), GL7 AF488 (GL7; BioLegend; 1:300), IgD PerCPCy5.5 (11- 26c2a; BD; 1:300), IgM BV421 (II/41; BD; 1:300), and B220 BV650 (RA3-6B2; BD; 1:300).
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