anti human igg4 fc  (SouthernBiotech)

Journal: Transfusion

Article Title: IgG4 anti‐ CD47 : Erythrophagocytosis and potential transfusion complications

doi: 10.1111/trf.70111

Figure Lengend Snippet: IgG4 antibody induced erythrophagocytosis and enhancement of alloantibody phagocytosis when CD47 is blocked. (A) IgG4 anti‐K (KEL1) causes erythrophagocytosis which is enhanced when CD47 is blocked using a deglycosylated anti‐CD47. (B) Polyclonal anti‐D (WinRho) erythrophagocytosis is enhanced in a dose‐dependent manner with deglycosylated anti‐CD47 blocking antibody. Results are using Student's t ‐test, mean ± SD for triplicates, using GraphPad. * p < .05, ** p < .01, *** p < .001, **** p < .0001.

Article Snippet: Drug or patients' diluted plasma was used to opsonize blood group O RBCs, washed, and stained with a PE‐labeled mouse monoclonal anti‐human IgG4 Fc (Southern Biotech, clone HP6025, Cat. # 9200‐09) for 30 min at 4°C.

Techniques: Blocking Assay

Journal: Transfusion

Article Title: IgG4 anti‐ CD47 : Erythrophagocytosis and potential transfusion complications

doi: 10.1111/trf.70111

Figure Lengend Snippet: Schematic depicting that (A) IgG4 anti‐CD47 leads to robust, largely FcγRI‐dependent erythrophagocytosis, but also involvement of FcRIIa but not FcRIIIa; and (B) antibodies to CD47 with functional Fc regions may block CD47 but contribute to phagocytosis and show an additive phagocytosis with unrelated alloantibodies. Ani‐CD47 with a nonfunctional Fc region may block CD47‐SIRPα (blocking the “don't eat me” signal) and lead to significant erythrophagocytosis of unrelated red blood cell (RBC) alloantibodies and may augment delayed hemolytic transfusion reactions (DHTRs).

Article Snippet: Drug or patients' diluted plasma was used to opsonize blood group O RBCs, washed, and stained with a PE‐labeled mouse monoclonal anti‐human IgG4 Fc (Southern Biotech, clone HP6025, Cat. # 9200‐09) for 30 min at 4°C.

Techniques: Functional Assay, Blocking Assay

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: IgG reactivity to (A) purified protein derivative (PPD) from Mtb , (B) Mtb cytosolic protein, (C) Mtb culture filtrate, (D) Ag85A and Ag85B, (E) Mtb cell wall, (F) ESAT-6 and CFP-10, and (G) control respiratory syncytial virus (RSV) was determined by customized multiplex Luminex for each individual patient sample with TB over three serial dilutions. Each dot represents relative reactivity for one individual patient sample determined by the area under the curve (AUC), summarizing the median fluorescence intensity (MFI) from serial dilutions. Bars represent median and 95% confidence intervals for latent ( n = 18) and active ( n = 19) TB groups. The p values determined by a Mann-Whitney U test are marked by ^ for significance after adjustment for multiple comparisons by Benjamini-Hochberg.

Article Snippet: Mouse Anti-Human IgG4 Fc-PE HP6025 , SouthernBiotech , Cat#9200-09; RRID:AB_2796693.

Techniques: Purification, Control, Virus, Multiplex Assay, Luminex, Fluorescence, MANN-WHITNEY

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) Representative chromatograms show patterns of individual glycoforms isolated from the Fc domain of antigen-specific and total bulk IgG in an individual patient with TB, determined by capillary electrophoresis. (B) Violin plots show the relative abundance of sialic acid, galactose, fucose, and bisecting N-acetylglucosamine (GlcNAc) across all individual glycoforms isolated from ESAT-6 and CFP-10 (red) and Mtb cell wall (blue) polyclonal IgG. Each dot represents an individual sample with latent ( n = 18) or active ( n = 19) TB. The median and interquartiles are shown. The dashed lines show the median RSV (green) and total bulk (purple) glycans. The p values determined by a Wilcoxon matched-pairs signed rank test are marked by ^ for significance after adjustment for multiple comparisons by Benjamini-Hochberg.

Article Snippet: Mouse Anti-Human IgG4 Fc-PE HP6025 , SouthernBiotech , Cat#9200-09; RRID:AB_2796693.

Techniques: Isolation, Electrophoresis

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) Luminescence from the virulent Mtb H37Rv luminescent reporter strain relates to colony-forming units (CFUs). The significance was evaluated by Pearson correlation. (B) To test the effect of antibodies on intracellular Mtb , primary human monocyte-derived macrophages were first infected with the virulent Mtb H37Rv luminescent reporter strain, and the extracellular bacteria were washed away. Then, Mtb -infected primary human monocyte-derived macrophages (MOI = 1) were treated with IgG. Finally, the bacterial burden was quantified with >99% of detectable Mtb in the intracellular as compared to the extracellular medium supernatant compartment. The error bars represent the mean ± SEM. Significance was determined by a Wilcoxon matched-pairs signed rank test. (C) Daily Mtb luminescence measurements representing the median of endemic control, active TB, and latent TB samples are shown for one representative healthy donor of human macrophages. (D) Data from n = 3 healthy human macrophage donors in independent experiments are summarized, with each dot representing the Mtb burden for each individual patient with TB relative to control polyclonal IgG. The median and 95% confidence interval (CI) are shown. The dashed line shows the median of endemic IGRA− control individuals. The significance was determined by a Mann-Whitney U test.

Article Snippet: Mouse Anti-Human IgG4 Fc-PE HP6025 , SouthernBiotech , Cat#9200-09; RRID:AB_2796693.

Techniques: Derivative Assay, Infection, Bacteria, Control, MANN-WHITNEY

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) The relationships between ESAT-6 and CFP-10 IgG levels and subclasses and intracellular Mtb burden within individuals with latent and active TB were evaluated by Spearman correlation. Heatmaps depict the Spearman rank correlation coefficient, ** p ≤ 0.01, and ^ stands for significance after adjustment for multiple comparisons by Benjamini-Hochberg. The scatterplot shows ESAT-6 and CFP-10 IgG1, and the Mtb burden is shown in the scatterplot, with each dot representing each individual with latent TB. (B) As a control, the relationships between Mtb cell-wall IgG levels and subclasses and intracellular Mtb burden are shown.

Article Snippet: Mouse Anti-Human IgG4 Fc-PE HP6025 , SouthernBiotech , Cat#9200-09; RRID:AB_2796693.

Techniques: Control

Journal: Cell reports

Article Title: ESAT-6 and CFP-10 reactive IgG in patients with tuberculosis inhibits intracellular bacteria

doi: 10.1016/j.celrep.2025.116653

Figure Lengend Snippet: (A) Each column in the histogram depicts the intracellular Mtb burden of one individual patient with latent (light gray) and active (dark gray) TB as in . The dashed line represents the intracellular Mtb burden with control IgG. (A and B) The anti- Mtb activity of IgG from each individual patient with TB was determined by the difference in Mtb burden between control and patient IgG (red). (C) For the n = 17 latent and n = 14 active TB samples with detectable anti- Mtb activities relative to ESAT-6 and CFP-10 IgG1, the relationships to ESAT-6 and CFP-10 IgG Fc glycans as determined by Spearman correlations are listed with ^ marking the significance after adjustment for multiple comparisons by Benjamini-Hochberg. (D) N-glycans from anti-ESAT-6 and CFP-10 mAb were enzymatically removed with PNGase F and then used to treat Mtb -infected primary human monocyte-derived macrophages. The intracellular Mtb burden is shown relative to a no-antibody (Ab) control. The graph summarizes data for n = 6 healthy macrophage donors, with each line representing a single donor. (E) An L234A and L235A (LALA) variant of anti-ESAT-6 and CFP-10 mAb was used to treat Mtb -infected primary human monocyte-derived macrophages. Each line represents a single healthy macrophage donor ( n = 9). The significance was determined by a Wilcoxon matched-pairs signed rank test.

Article Snippet: Mouse Anti-Human IgG4 Fc-PE HP6025 , SouthernBiotech , Cat#9200-09; RRID:AB_2796693.

Techniques: Control, Activity Assay, Infection, Derivative Assay, Variant Assay

Journal: EMBO Molecular Medicine

Article Title: Dynamics of checkpoint receptors in γδ T cell subsets are associated with clinical response during anti-PD-1 immunotherapies

doi: 10.1038/s44321-025-00338-9

Figure Lengend Snippet: Analysis of PD-1 expression in PBMCs from patients with stage IV metastatic melanoma who either responded (R, n = 9) or did not respond (NR, n = 8) to pembrolizumab (therapeutic α-PD-1) treatment. Paired samples were obtained from these patients before (pre) and 3 to 4 months after (post) the start of immunotherapy. ( A ) To assess total PD-1 expression in these samples, we used two fluorescently labeled antibodies (left), targeting either PD-1 molecules (α-PD-1-BUV-737) or the therapeutic α-PD1 antibody (α-IgG4-PE). The total frequency of PD-1 + cells was determined as cells that were positive for α-PD-1-BUV-737 and α-IgG4-PE; the frequency of therapeutic antibody binding (thera PD-1 + ) was determined as cells that were positive only for α-IgG4-PE. ( B ) Representative flow cytometric analysis of therapeutic α-PD-1 antibody binding on the surface of Vδ1 and Vδ2 cells. ( C ) Days between the last antibody infusion and the collection of samples. Each symbol represents a patient. ( D ) Thera PD-1 + Vδ1 and Vδ2 cells after pembrolizumab treatment. P values were calculated by paired two-way ANOVA and Tukey’s multiple comparisons test. Each symbol represents a patient. ( E ) Frequency of Vδ1 cells with bound therapeutic antibody out of the total PD-1 + Vδ1 population after pembrolizumab treatment. The pie charts show average expression in 9 R and 8 NR patients. ( F ) Thera PD-1 + CD4 and CD8 T cells after pembrolizumab treatment. Each symbol represents a patient. ( G ) Progression-free survival (PFS; β = −2.5, P = 0.021, HR = 0.083 with 95% CI of 0.01–0.68), and overall survival (OS; β = −1.8, P = 0.03, HR = 0.16 with 95% CI of 0.03–0.84) curves according to Cox proportional hazard model of patients with melanoma based on frequency of thera PD-1 + Vδ1 (cut-off >14%). ( H ) Progression-free survival (PFS; β = −1.1, P = 0.057, HR = 0.32 with 95% CI of 0.099–1), and overall survival (OS; β = −1.1, P = 0.1, HR = 0.34 with 95% CI of 0.095–1.2) curves according to Cox proportional hazard model of patients with melanoma based on frequency of thera PD-1 + CD8 (cut-off >5.5%). ( G , H ) The cut-off was calculated as the average between the median frequency of thera PD-1 + cells in patients who responded to therapy and the median frequency of thera PD-1 + cells in the patients who did not respond to therapy. .

Article Snippet: Anti-human IgG4 PE , Southern Biotech , 9200-09.

Techniques: Expressing, Labeling, Binding Assay