hrp conjugated goat anti mouse igg2a  (SouthernBiotech)

Journal: bioRxiv

Article Title: Dirty mice better recapitulate key features of mRNA vaccine immunogenicity observed in humans

doi: 10.64898/2026.03.07.709392

Figure Lengend Snippet: mRNA vaccination in dirty mice requires booster vaccination and is less durable than in SPF mice. (A) Model for generating and vaccinating dirty mice, days (d) indicated. (B–E) Serum antibody responses in SPF (black) and dirty (red) mice. Illustration generated with images from NIAID NIH BioArt. (B) Anti-S1 RBD IgG levels over 61 days post-vaccination (N=48), expressed as area under the curve (AUC) from serial dilution ELISAs (OD 405 ). (C) Quantification of serum IgG specific to SARS-CoV-2 WT Spike (AU/ml) through day 151 post-immunization. (D) Neutralizing antibody titers over 61 days post-vaccination, expressed as FRNT 50 . (E) Cross-reactive serum IgG levels against SARS-CoV-2 Spike variants (WT, BA.1, BA.5) at day 61 (AU/ml). Data represent individual mice with mean ± SEM; p-values determined by Welch’s T-test 2 . (F) Antibody clearance from serum of mock-injected (N=3) and α’NP-injected SPF (N=4) and dirty mice (N=4), expressed as α’NP IgG2a AUC. N=3-4 AUC values per timepoint are represented on the graph aside from mock d59 with N=2, with standard deviation indicated. Antibody levels in B, C, and E were determined using MSD-ECLIA. Antibody levels in F were determined with ELISA.

Article Snippet: Serum was diluted 1:100 in dilution buffer prior to serial 1:4 dilutions, and Goat Anti-Mouse IgG2a Human ads-HRP secondary antibody (SouthernBiotech) was used as the secondary antibody for detection.

Techniques: Generated, Serial Dilution, Injection, Standard Deviation, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Dirty mice better recapitulate key features of mRNA vaccine immunogenicity observed in humans

doi: 10.64898/2026.03.07.709392

Figure Lengend Snippet: The dirty mouse model is stable over time and season. (A) Percent of mice in each cohoused cohort that were seropositive for murine pathogens. Data corresponds to mice shown in . Only pathogens with seropositivity among the mice are depicted, additional pathogens tested in panel are listed in Materials and Methods. (B) Correlation of T cell activation status (percent of CD44+-hi expressing cells among CD8a+ T cells) with anti-S1 RBD IgG levels at 60 days post-prime (30 days post-boost). Antibody levels are represented as AUC. N=21 mice. Linear regression model equation and 95% confidence interval is shown on graph. (C) Multidimensional scaling (MDS) analysis of serology panel data from cohoused mice. Each dot represents a dirty laboratory mouse. Distance between points represents similarity of pathogen exposure history. Points are colored by the year that the experiment occurred. Data is representative of N=1014 mice. (D) Comparison of average percent CD44+-Hi among CD8a+ T cells (left) and average pathogen richness (right) from mice in panel C across seasons. Error bars represent standard deviation. (E) Experimental design used to generate the double co-housed dirty mouse model. Illustration generated with images from NIAID NIH BioArt. (F) Heatmaps of seroprevalence against a panel of murine pathogens in single co-housed (60 days) and double co-housed (120 days) mice. Squares are divided into triangles representing data from two independent experimental replicates. (G) Frequency of activated CD8a+/CD44-Hi T cells in single and double co-housed mice assessed at days 60 and 121. Bars represent mean ± SEM. (H) Anti-S1 RBD IgG log10 endpoint titers (OD 405 ) 30 days post-prime and -booster (N=20) in SPF (black), single (red), and double (purple) co-housed mice. Data represent individual mice with horizontal bars indicating mean and brackets indicate fold-change relative to SPF. Antibody levels in panels B and H were determined using ELISA.

Article Snippet: Serum was diluted 1:100 in dilution buffer prior to serial 1:4 dilutions, and Goat Anti-Mouse IgG2a Human ads-HRP secondary antibody (SouthernBiotech) was used as the secondary antibody for detection.

Techniques: Activation Assay, Expressing, Comparison, Standard Deviation, Generated, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Experimental timeline of the immunisation (i.e., vaccination) against OVA and subsequent tumour inoculation of the OVA-expressing B16F10 cell lines in mice. B) Titers of anti-OVA IgG per IgG subtypes in the plasma of the mice at d18. C) Percentage of OVA-specific CD8 + T cells against the immunogenic epitope OVA 257-264 (SIINFEKL) in the blood at d18 in pre-immunised mice (vax) or naïve mice (no vax) (n ≥ 3, mean ± SD, unpaired t-test). D, E) B16F10 melanoma cells, either wild-type (WT) or genetically modified to express high ( HI ) or low ( LO ) doses of membrane-bound OVA (B16mOVA) or soluble OVA (B16-OVA), were injected intradermally in C57BL/6 mice. Tumour growth (D) and associated survival (E) of the different OVA-expressing B16 cell lines in pre-immunised mice (n ≥ 5, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d10, d14 or d26 for tumour growth, log-rank tests for survival). F) Potential anti-tumour mechanisms engaged by soluble (left) or membrane-bound (right) xenoantigens when expressed by cancer cells.

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Expressing, Clinical Proteomics, Genetically Modified, Membrane, Injection

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Immunofluorescent staining of B16F10 WT melanoma with the TRP1-targeting TA99 antibody (scale bar = 100 μm). B) B16F10 WT tumour growth upon treatment with TA99 or IgG2a isotype control (n ≥ 4, mean ± SEM, Mann-Whitney test on d11). C) Design of FabTRP-OVA fusion protein and analysis of its production by SDS-PAGE (expected size = 92,3 kDa). D) Illustration of the FabTRP-OVA fusion protein approach for targeting the xenoantigen to the cancer cell membrane. E) Representative flow cytometry plot of FabTRP-OVA, OVA and no OVA (i.e., secondary antibody only) binding to the surface of B16F10 WT cells. F) B16F10 WT tumour growth when treated with FabTRP-OVA in pre-immunised or naïve mice (n ≥ 4, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d12). G) B16F10 WT tumour growth when treated with FabTRP-OVA in combination with anti-PD-1 checkpoint blocade therapy (n ≥ 4, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d12). H) Experimental treatment timeline of xenoantigen delivery (i.e., FabTRP-OVA, OVA or PBS (vehicle only)) and anti-PD-1 checkpoint blockade therapy in B16F10 WT in mice pre-immunised against OVA (Vax) or naïve (No vax). I, J) B16F10 WT tumour growth (I) and associated mouse survival (J) upon treatment with FabTRP-OVA, OVA or PBS in combination with anti-PD-1 checkpoint blockade therapy (n ≥ 5, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d16, log-rank tests for survival).

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Staining, Control, MANN-WHITNEY, SDS Page, Membrane, Flow Cytometry, Binding Assay

Journal: bioRxiv

Article Title: Membrane localisation and checkpoint blockade enhance xenoantigen delivery to redirect pre-existing immunity against tumours

doi: 10.64898/2026.03.01.708859

Figure Lengend Snippet: A) Experimental timeline of the immunisation (i.e., vaccination) against varicella VZVO using Varivax or a combination of gE/CpG. B) IFΝγ quantification in the supernatant of gE restimulated (+) or non-restimulated (-) splenocytes, collected from mice immunized with Varivax, gE/CpG or PBS (i.e., naïve). Ionomycin+PMA was used as a positive control. C) Titers of anti-gE total IgG and per IgG subtypes in the plasma of the pre-immunised mice at d55 (before tumour implantation). D) Experimental treatment timeline of xenoantigen delivery (i.e., Varivax, gE or PBS (vehicle only)) and anti-PD-1 checkpoint blockade treatment in B16F10 WT in mice pre-immunised with Varivax against varicella VZVO . E, F) B16F10 WT tumour growth (E) and associated mouse survival (F) upon treatment with Varivax, gE or PBS in combination with anti-PD-1 checkpoint blockade (n ≥ 7, mean ± SEM, Kruskal-Wallis with Dunn’s post-test on d16, log-rank tests for survival).

Article Snippet: Samples were applied to the antigen coated plate and incubated at RT for 2 h. The plates were washed three times with PBST and HRP-conjugated antibodies were used to detect antigen-specific antibodies: anti-mouse IgG1 (#1070-05), anti-mouse IgG2a (#1080-05), anti-mouse IgG2b (#1090-05) and anti-mouse IgG3 (#1100-05) from Southern Biotech (Birmingham, AL, USA).

Techniques: Positive Control, Clinical Proteomics