Membrane:Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice
Article Snippet: The FluoroSpot assay was performed using mouse IFN-γ/TNF-α/IL-2 Triple-Color FluoroSpot kits (Cellular Technology Limited (CTL), Cleveland, OH). .. A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ). .. A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-Test medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).
Article Title: CoVaccine HT™ Adjuvant Potentiates Robust Immune Responses to Recombinant SARS-CoV-2 Spike S1 Immunization
Article Snippet: .. A total of 2.5 x 10 5 cells per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added to a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 40 h with peptides, PepTivator ® SARS-CoV-2 Prot_S1 peptide pool consisting of 15-mer peptides with 11 amino acids overlapping, covering the N-terminal S1 domain of the Spike protein of SARS-CoV-2 (Miltenyi Biotec, Auburn, CA) at 0.2 μg/ml and 0.5 μg/ml per peptide, or medium alone. .. Splenocytes at (5 x 10 4 per well) were incubated with PMA (0.01 μM)/Ionomycin (0.167 μM) cocktail (BioLegend, San Diego, CA) as a positive control.
Article Title: CoVaccine HT™ adjuvant potentiates robust immune responses to recombinant SARS-CoV-2 Spike S1 immunisation
Article Snippet: .. A total of 2.5 × 10 cells per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added to a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 40 hours with peptides, PepTivator ® SARS-CoV-2 Prot_S1 peptide pool consisting of 15-mer peptides with 11 amino acids overlapping, covering the N-terminal S1 domain of the Spike protein of SARS-CoV-2 (Miltenyi Biotec, Auburn, CA) at 0.2 μg/mL and 0.5 μg/mL per peptide, or medium alone. .. Splenocytes at (5 × 10 4 per well) were incubated with PMA (0.01 μM) /Ionomycin (0.167 μM) cocktail (BioLegend, San Diego, CA) as a positive control.
Bioprocessing:Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice
Article Snippet: The FluoroSpot assay was performed using mouse IFN-γ/TNF-α/IL-2 Triple-Color FluoroSpot kits (Cellular Technology Limited (CTL), Cleveland, OH). .. A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ). .. A total of 3.0 x 10 5 splenocytes per well in serum-free CTL-Test medium (Cellular Technology Limited, Shaker Heights, OH) were added in a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 24 hours at 37°C, 5% CO2 with pools of 15-mer peptides with 11 overlapping amino acids (5 μg/mL per peptide), covering the entire LASV GP (GenScript, Piscataway, NJ).
Article Title: CoVaccine HT™ Adjuvant Potentiates Robust Immune Responses to Recombinant SARS-CoV-2 Spike S1 Immunization
Article Snippet: .. A total of 2.5 x 10 5 cells per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added to a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 40 h with peptides, PepTivator ® SARS-CoV-2 Prot_S1 peptide pool consisting of 15-mer peptides with 11 amino acids overlapping, covering the N-terminal S1 domain of the Spike protein of SARS-CoV-2 (Miltenyi Biotec, Auburn, CA) at 0.2 μg/ml and 0.5 μg/ml per peptide, or medium alone. .. Splenocytes at (5 x 10 4 per well) were incubated with PMA (0.01 μM)/Ionomycin (0.167 μM) cocktail (BioLegend, San Diego, CA) as a positive control.
Article Title: CoVaccine HT™ adjuvant potentiates robust immune responses to recombinant SARS-CoV-2 Spike S1 immunisation
Article Snippet: .. A total of 2.5 × 10 cells per well in serum-free CTL-TestTM medium (Cellular Technology Limited, Shaker Heights, OH) were added to a 96 well PVDF membrane plate pre-coated with capture monoclonal antibodies and stimulated for 40 hours with peptides, PepTivator ® SARS-CoV-2 Prot_S1 peptide pool consisting of 15-mer peptides with 11 amino acids overlapping, covering the N-terminal S1 domain of the Spike protein of SARS-CoV-2 (Miltenyi Biotec, Auburn, CA) at 0.2 μg/mL and 0.5 μg/mL per peptide, or medium alone. .. Splenocytes at (5 × 10 4 per well) were incubated with PMA (0.01 μM) /Ionomycin (0.167 μM) cocktail (BioLegend, San Diego, CA) as a positive control.
Enzyme-linked Immunospot:Article Title: Split tolerance permits safe Ad5-GUCY2C-PADRE vaccine-induced T-cell responses in colon cancer patients
Article Snippet: Human IFNγ SC Enzymatic ELISpot plates (Cellular Technology Limited) were coated according to the manufacturer’s instructions. .. PBMC samples were thawed using CTL Anti-Aggregate WashTM (Cellular Technology Limited) according to the manufacturer’s instructions and 5 × 10 5 viable cells/well were plated in ELISpot plates in serum-free CTL-TestTM Medium (Cellular Technology Limited) without a rest period. .. Antigens were prepared and added to PBMCs at the indicated final concentrations in CTL-TestTM Medium (Cellular Technology Limited) with 1% DMSO in all conditions: 1% DMSO (ATCC); 1 μg/mL Ad5 Peptide Mix (PM-HAdV5, JPT Peptide Technologies, Berlin, Germany); 2 and 10 μg/mL human GUCY2C Peptide Mix (custom 15 mer/11 aa overlap library of human GUCY2C 1–429 , JPT Peptide Technologies); 1 μg/mL PADRE (BAP-251, EMC Microcollections, Tübingen, Germany).
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