anti digoxin fitc  (Vector Laboratories)

Journal: Vaccines

Article Title: The Long-Term Immunity of a Microneedle Array Patch of a SARS-CoV-2 S1 Protein Subunit Vaccine Irradiated by Gamma Rays in Mice

doi: 10.3390/vaccines13010086

Figure Lengend Snippet: A comparison of immunogenicity in mice immunized with four SARS-CoV-2 rS1Delta protein subunit vaccines. ( A ) Schedule of immunization and blood sampling for IgG end point titration. Balb/c mice (5 per group) were immunized with 45 μg of either rS1, rS1RS09, rS1f, or rS1fRS09 proteins of SARS-CoV-2 Delta then administered intramuscularly at week 0 and 3. Syringes indicated the timing of immunizations, and the red drops denote times at which blood was drawn. ( B ) Sera were diluted, and SARS-CoV-2-S1-specific antibodies were quantified by ELISA to determine the IgG endpoint titer. The IgG titers at each time points were showed in each mouse. The bars represent the geometric mean with geometric SD. ( C , D ) Sera at weeks 0, 5, and 18 were diluted, and SARS-CoV-2-S1WU-specific IgG1 ( C ) and IgG2a ( D ) were quantified by ELISA to determine each IgG subclasses’ endpoint titer. The titers at each time points were showed for each mouse. The bars represent the geometric mean. ( E ) S1-specific IgG1/IgG2a ratios of individual mice at weeks 7 and 18 as mean values with SEM. ( F ) Flow cytometry assay of Expi293 cells expressing S1Delta-S2WU at the cell surface. At 36 h post-transfection with pAd/S1Delta-S2WU, the binding to SARS-CoV-2-S at the Expi293 cell surface was analyzed by incubation with mice sera obtained at week 7 after immunization followed by staining with FITC-conjugated anti-mouse IgG. Groups were compared by the Kruskal–Wallis test at each time point, followed by Dunn’s multiple comparisons. Significant differences are indicated by * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Half a million cells were incubated with 1 μL of the mouse serum from each group or 1 μL of the monoclonal antibody (D003, Sino Biological) for 30 min, followed by staining with a FITC-conjugated anti-mouse secondary IgG antibody (Jackson ImmunoResearch).

Techniques: Comparison, Immunopeptidomics, Vaccines, Sampling, Titration, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Transfection, Binding Assay, Incubation, Staining

Journal: Vaccines

Article Title: The Long-Term Immunity of a Microneedle Array Patch of a SARS-CoV-2 S1 Protein Subunit Vaccine Irradiated by Gamma Rays in Mice

doi: 10.3390/vaccines13010086

Figure Lengend Snippet: Immune responses induced by different vaccine routes of the administration of high doses. ( A ) Silver stained SDS-PAGE of purified SARS-CoV-2 Delta rS1RS09 proteins. ( B ) Fabrication of dissolving microneedle patch; MAP-rS1RS09 was composed of 35 microneedles. ( C ) Each MN was approximately 700 μm in length, occupying a 2.36 cm 2 area size of the patch, 7 × 5 arrays of 700 μm-long microneedles with 45 μg of rS1RS09Delta. ( D ) Experimental schedule representing the immunization timeline. Balb/c mice (5 per group) were primed and boosted at three week intervals with either 45 μg of rS1RS09Delta intramuscularly or MAP-rS1RS09Delta intracutaneously. The red drops represented bleeding. ( E ) Reciprocal serum endpoint dilutions of SARS-CoV-2-S1-specific antibodies were measured by ELISA to determine the IgG endpoint titers until week 90. Sera at weeks 0, 5, and 18 were diluted, and SARS-CoV-2-S1WU-specific IgG1 ( F ) an IgG2a ( G ) were quantified by ELISA to determine each IgG subclasses’ endpoint titer. The titers at each of the time points were showed for each mouse. The bars represent the geometric mean. ( H ) S1-specific IgG1/IgG2a ratios of individual mice at weeks 7 and 18 as mean values with SEM. IM and MAP groups were compared for statistically significant differences using a non-parametric Mann–Whitney U test compared with pre-immunized sera. * p < 0.05, ** p < 0.01, n.s., not significant.

Article Snippet: Half a million cells were incubated with 1 μL of the mouse serum from each group or 1 μL of the monoclonal antibody (D003, Sino Biological) for 30 min, followed by staining with a FITC-conjugated anti-mouse secondary IgG antibody (Jackson ImmunoResearch).

Techniques: Staining, SDS Page, Purification, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY