Journal: NPJ Vaccines

Article Title: Deimmunization of flagellin for repeated administration as a vaccine adjuvant

doi: 10.1038/s41541-021-00379-4

Figure Lengend Snippet: a Characterization of dFlaB and FlaB ΔD2D3 by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), native-PAGE, and subsequent Western blot analysis with mouse anti-FlaB serum induced by intraperitoneal immunization of FlaB formulated complete Freund’s adjuvant. b Determination of TLR5-dependent NF-κB stimulating activity by FlaB, dFlaB, and FlaB ΔD2D3 . The relative NF-κB activities were analyzed by using HEK-Blue TM hTLR5 cells and HEK-Blue TM detection assay systems. EC50 were calculated using triplicate OD 620 nm values for each protein concentration over a wide range of protein concentrations (0.03719 nM to 76.16 nM at the AAT Bioquest website ( https://www.aatbio.com/tools/ec50-calculator ). The same molar ratio of proteins was used, and PBS was used as a negative control.

Article Snippet: We measured TLR5-dependent NFκB stimulating activity of the recombinant proteins by using HEK-Blue TM hTLR5 cells (InvivoGen, hκb-htlr-5) and HEK-Blue TM Detection (InvivoGen, hb-det2) assay systems following the manufacturer’s instruction.

Techniques: Polyacrylamide Gel Electrophoresis, SDS Page, Clear Native PAGE, Western Blot, Adjuvant, Activity Assay, Detection Assay, Protein Concentration, Negative Control