other:Article Title: High Frequency of Prior Severe Acute Respiratory Syndrome Coronavirus 2 Infection by Sensitive Nucleocapsid Assays
Article Snippet: Cells were washed twice with 1× Perm Wash buffer (BD Perm/Wash Buffer 10× in the CytoFix/CytoPerm fixation/permeabilization kit diluted with MilliQ water and passed through 0.22 μm filter) and stained intracellularly with monoclonal antibodies against Ki67 (clone B56, BB515), interleukin (IL) 21 (clone 3A3-N2.1, PE), CD69 (clone TP1.55.3, ECD), IL-10 (clone JES3-9D7, PE CY7), IL-13 (clone JES10-5A2, BV421), IL-4 (clone MP4-25D2, BV605), tumor necrosis factor–α (clone Mab11, BV650), IL-17 (clone N49-653, BV750), interferon gamma (IFN-γ) (clone B27; BUV395), IL-2 (clone MQ1-17H12, BUV737), IL-6 (clone MQ2-13A5, APC), and CD3 (clone SP34.2, Alexa 700) for 30 minutes.
Article Title: Early Emerging SARS-CoV-2 Spike Mutants Are Diversified in Virologic Properties but Elicit Compromised Antibody Responses.
Article Snippet: Cells were then washed twice with 1x Perm Wash buffer (BD Perm/WashTM Buffer 10× in CytoFix/CytoPerm Fixation/Permeabilization kit diluted with MilliQ water and passed through 0.22 μm filter) and stained intracellularly with mAbs against IFN-γ (clone XMG1.2; BV421), IL-2 (clone JES6-5H4; PE), IL-4 (clone 11B11; APC), and TNFα (clone MP6-XT22; Alexa Fluor 700) (all antibodies diluted 1:100) for 1 h. Cells were then washed twice with 1× Perm Wash buffer and washed once with MACS buffer, then fixed with 250 μL of freshly prepared 2% formaldehyde.
Article Title: SARS-CoV-2 XBB.1.5 mRNA booster vaccination elicits limited mucosal immunity.
Article Snippet: Cells were washed twice with a 1× Perm Wash buffer (BD Perm/Wash Buffer 10× in the CytoFix/CytoPerm Fixation/Permeabilization kit diluted with MilliQ water and passed through a 0.22- μm filter) and stained intracellularly with monoclonal antibodies against Ki67 (clone B56, BB515; 0.25 μl per test), interleukin- 21 (IL- 21) [clone 3A3- N2.1, phycoerythrin (PE); 5 μl per test], CD69 (clone TP1.55.3, ECD; 0.75 μl per test), IL- 10 (clone JES3- 9D7, PE- CY7; 2 μl per test), IL- 13 (clone JES10- 5A2, BV421; 2.5 μl per test), IL- 4 (clone MP4- 25D2, BV605; 2.5 μl per test), TNF- α (tumor necrosis factor–α) (clone Mab11, BV650; 2.5 μl per test), IL- 17 (clone N49- 653, BV750; 2 μl per test), IFN- γ (clone B27; BUV395; 2.5 μl per test), IL- 2 (clone MQ1- 17H12, BUV737; 2.5 μl per test), IL- 6 (clone MQ2- 13A5, allophycocyanin; 1.25 μl per test), and CD3 (clone SP34.2, Alexa Fluor 700; 0.25 μl per test) for 30 min.
Article Title: Protection against SARS-CoV-2 Omicron BA.1 variant challenge in macaques by prime-boost vaccination with Ad26.COV2.S and SpFN.
Article Snippet: Jingyou Yu1†, Paul V. Thomas2,3†, Katherine McMahan1, Catherine Jacob-Dolan1,4,5, Jinyan Liu1, Xuan He1, David Hope1, Elizabeth J. Martinez2,3, Wei-Hung Chen2,3, Michaela Sciacca1, Nicole P. Hachmann1, Michelle Lifton1, Jessica Miller1, Olivia C. Powers1, Kevin Hall1, Cindy Wu1, Julia Barrett1, Isabella Swafford2,3, Jeffrey R. Currier2,3, Jocelyn King6, Courtney Corbitt2,3, William C. Chang2, Emily Golub2,3, Phyllis A. Rees2,3, Caroline E. Peterson2,3, Agnes Hajduczki2,3, Elizabeth Hussin3,7, Camille Lange3,7, Hua Gong3,7, Gary R. Matyas7, Mangala Rao7, Dominic Paquin-Proulx2,3, Gregory D. Gromowski6, Mark G. Lewis8, Hanne Andersen8, Meredith Davis-Gardner9, Mehul S. Suthar9, Nelson L. Michael10, Diane L. Bolton2,3, M. Gordon Joyce2,3*, Kayvon Modjarrad2, Dan H. Barouch1,4,5*
Article Title: High Frequency of Prior SARS-CoV-2 Infection by Sensitive Nucleocapsid Assays.
Article Snippet: Cells were washed twice with 1× Perm Wash buffer (BD Perm/Wash Buffer 10× in the CytoFix/CytoPerm fixation/ permeabilization kit diluted with MilliQ water and passed through 0.22 μm filter) and stained intracellularly with monoclonal antibodies against Ki67 (clone B56, BB515), interleukin (IL) 21 (clone 3A3-N2.1, PE), CD69 (clone TP1.55.3, ECD), IL-10 (clone JES3-9D7, PE CY7), IL-13 (clone JES10-5A2, BV421), IL-4 (clone MP4-25D2, BV605), tumor necrosis factor–α (clone Mab11, BV650), IL-17 (clone N49-653, BV750), interferon gamma (IFN-γ) (clone B27; BUV395), IL-2 (clone MQ1-17H12, BUV737), IL-6 (clone MQ2-13A5, APC), and CD3 (clone SP34.2, Alexa 700) for 30 minutes.
Article Title: Immunogen characterization reveals an intrinsic hindrance in eliciting neutralizing antibodies against JN.1 variant
Article Snippet: Cells were then washed twice with 1 × Perm Wash buffer (BD Perm/WashTM Buffer 10× in CytoFix/CytoPerm Fixation/Permeabilization kit diluted with MACS buffer and passed through 0.22 μm filter) and stained intracellularly with mAbs against IFN-γ (clone XMG1.2; BV421), IL-2 (clone JES6-5H4; PE), IL-4 (clone 11B11; APC), and TNFα (clone MP6-XT22; Alexa Fluor 700) (all antibodies diluted 1:100) for 1 h. Cells were then washed twice with 1× Perm Wash buffer and washed once with MACS buffer, then fixed with 250 μL of freshly prepared 2% formaldehyde.
Article Title: Waning immunity and IgG4 responses following bivalent mRNA boosting.
Article Snippet: Cells were washed twice with 1× Perm Wash buffer (BD Perm/Wash buffer 10× in the CytoFix/CytoPerm Fixation/Permeabilization kit diluted with MilliQ water and passed through 0.22- μm filter) and stained intracellularly with mAbs against Ki67 (clone B56, BB515), interleukin- 21 (IL- 21) [clone 3A3- N2.1, phycoerythrin (PE)], CD69 [clone TP1.55.3, extracellular domain (ECD)], IL- 10 (clone JES3- 9D7, PE CY7), IL- 13 (clone JES10- 5A2, BV421), IL- 4 (clone MP4- 25D2, BV605), tumor necrosis factor–α (TNF- α) (clone Mab11, BV650), IL- 17 (clone N49653, BV750), IFN- γ (clone B27; BUV395), IL- 2 (clone MQ1- 17H12, BUV737), IL- 6 (clone MQ2- 13A5, APC), and CD3 (clone SP34.2, Alexa Fluor 700) for 30 min.
Staining:Article Title: Immunogenicity and Efficacy of TNX-1800, A Live Virus Recombinant Poxvirus Vaccine Candidate, against SARS-CoV-2 Challenge in Nonhuman Primates.
Article Snippet: The cells were subsequently washed twice with 2% FBS in DPBS buffer and incubated for 15 min with 200 uL of BD Cytofix/CytoPerm Fixation/Permeabilization solution. .. The cells were washed twice with 1× Perm Wash buffer (BD Perm Wash Buffer 10× in the Cytofix/CytoPerm Fixation Permeabilization kit diluted with MilliQ water and passed through a 0.22 um filter) and stained intracellularly with monoclonal antibodies against IFN-γ and IL-4 for 30 min. .. The fixed cells were transferred to 96-well round bottom plates and analyzed by the Cytek Aurora flow cytometer (Cytek Biosciences, Freemont, CA, USA).
Bioprocessing:Article Title: Immunogenicity and Efficacy of TNX-1800, A Live Virus Recombinant Poxvirus Vaccine Candidate, against SARS-CoV-2 Challenge in Nonhuman Primates.
Article Snippet: The cells were subsequently washed twice with 2% FBS in DPBS buffer and incubated for 15 min with 200 uL of BD Cytofix/CytoPerm Fixation/Permeabilization solution. .. The cells were washed twice with 1× Perm Wash buffer (BD Perm Wash Buffer 10× in the Cytofix/CytoPerm Fixation Permeabilization kit diluted with MilliQ water and passed through a 0.22 um filter) and stained intracellularly with monoclonal antibodies against IFN-γ and IL-4 for 30 min. .. The fixed cells were transferred to 96-well round bottom plates and analyzed by the Cytek Aurora flow cytometer (Cytek Biosciences, Freemont, CA, USA).
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