igg2  (SouthernBiotech)

Journal: medRxiv

Article Title: Anti-Spike IgG4 and Fc Effector Responses: The Impact of SARS-CoV-2 Vaccine Platform–Specific Priming and Immune Imprinting

doi: 10.1101/2025.03.19.25324267

Figure Lengend Snippet:

Article Snippet: The plates were again washed with PBST, and then a secondary antibody: mouse anti-human IgG1 (Sino Biological), a mouse anti-human IgG2 (SouthernBiotech), a mouse anti-human IgG3, (Sino Biological) a mouse anti-human IgG4 (Invitrogen) conjugated with horseradish peroxidase was added and incubated for 1 h at room temperature (RT).

Techniques:

Journal: medRxiv

Article Title: Anti-Spike IgG4 and Fc Effector Responses: The Impact of SARS-CoV-2 Vaccine Platform–Specific Priming and Immune Imprinting

doi: 10.1101/2025.03.19.25324267

Figure Lengend Snippet: Anti-Spike (rS) IgG isotypes in subjects primed with either with mRNA vaccines (mRNA-1273 or BNT162b2) or the protein-based subunit COVID-19 vaccine (NVX-CoV2373). Pre-booster (day 0), post-booster NVX-CoV2373 (day 28) or mRNA (BNT162b2) vaccines (day 29). A. Schematic of the vaccines administered to subjects. ELISA-based quantitation [Geometric Mean Titers (GMT) with 95% confidence intervals (CI)] of IgG isotypes is shown in B–E : Gray – mRNA vaccines x3 +NVX-CoV2373 (N=19), dark-blue – NVX-CoV273 x3 + NVX-CoV273 (N=18), F–I: Light-blue – NVX-CoV273 x2 + BNT162b2 (N=24 for Day 0 and N=30 for Day 29). Statistical analysis was performed by the two-tailed Mann–Whittney test. **p<0.01, ***p < 0.001, ****p < 0.0001.n.s. indicates not significant.

Article Snippet: The plates were again washed with PBST, and then a secondary antibody: mouse anti-human IgG1 (Sino Biological), a mouse anti-human IgG2 (SouthernBiotech), a mouse anti-human IgG3, (Sino Biological) a mouse anti-human IgG4 (Invitrogen) conjugated with horseradish peroxidase was added and incubated for 1 h at room temperature (RT).

Techniques: Vaccines, Enzyme-linked Immunosorbent Assay, Quantitation Assay, Two Tailed Test

Journal: medRxiv

Article Title: Anti-Spike IgG4 and Fc Effector Responses: The Impact of SARS-CoV-2 Vaccine Platform–Specific Priming and Immune Imprinting

doi: 10.1101/2025.03.19.25324267

Figure Lengend Snippet: Anti-Spike (rS) IgG isotype proportions in subjects receiving the protein-based subunit XBB.1.5 COVID-19 vaccine (NVX-CoV2601) after priming with mRNA vaccines (≥3x). Pre-booster (day 0), post-booster with the NVX-CoV2601 vaccine (day 28). A. Anti-rS IgG isotype proportions (pie chart) against the wild-type (WT) trimer antigen (ancestral) B. Anti-rS IgG isotype proportions (pie chart) against the XBB.1.5 trimer antigen C. Anti-rS IgG isotype levels (GMT with 95% CI bar graph) against WT (Stripes, N=29) and XBB.1.5 (Solid color, N=29) trimer antigens. Geometric mean fold raise (GMFR) at day 28 post booster compared to pre-booster levels (day 0) are shown above the bars. Light-gray – anti-rS IgG1, medium-gray – anti-rS IgG2, black – anti-rS IgG3, plum – anti rS IgG4. Statistical analysis was performed by the two-tailed Mann–Whittney test. **p < 0.01, ***p<0.001 indicates significance. n.s. indicates not significant.

Article Snippet: The plates were again washed with PBST, and then a secondary antibody: mouse anti-human IgG1 (Sino Biological), a mouse anti-human IgG2 (SouthernBiotech), a mouse anti-human IgG3, (Sino Biological) a mouse anti-human IgG4 (Invitrogen) conjugated with horseradish peroxidase was added and incubated for 1 h at room temperature (RT).

Techniques: Vaccines, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Spike specific IgG3 and nucleocapsid IgG response in serum serve as distinguishing immunological markers between SARS-CoV-2 infection and vaccination

doi: 10.3389/fimmu.2025.1518915

Figure Lengend Snippet: RBD specific IgG and its subclasses after SARS-CoV-2 natural infection. RBD specific IgG (A) and RBD specific IgG1 (B) , IgG2 (C) , IgG3 (D) , IgG4 (E) was analyzed in serum samples from naturally infected COVID-19 patients (n=39) and healthy controls (n=20). Samples were collected at different time points are indicated below the graph. Each symbol represents one individual where different color of dots indicates the category of patients as asymptomatic (black), mild (green), moderate (blue) and severe (orange). Statistical analysis was performed between healthy controls and patients using the Man-Whitney test. *P <0.05, ***P <0.0001, ns, not significant.

Article Snippet: Plates were then loaded with mouse anti-human Fc specific IgG1, IgG2, IgG3 and IgG4 that were conjugated with HRP (Southern Biotech) and incubated for another hour at room temperature (RT).

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Spike specific IgG3 and nucleocapsid IgG response in serum serve as distinguishing immunological markers between SARS-CoV-2 infection and vaccination

doi: 10.3389/fimmu.2025.1518915

Figure Lengend Snippet: Comparison between RBD specific IgG1 and IgG3 after natural infection and vaccination. RBD-specific IgG1 and IgG3 antibody titers (mean +SEM) are shown in graph (A) after COVID-19 infection at Day-1, Day-7 and Day-28. RBD-specific IgG1 and IgG3 antibodies were compared at 1 month (post dose-1), 2/3 Months (post dose-2) and 6 months (Post 6M) after the first vaccination shown in graph (B) . Each dark bar represents mean of IgG3 with SEM whereas white bar represents IgG1. Statistical analyses were performed between IgG1 and IgG3 using the Wilcoxon test. **P <0.001, ***P <0.0001, ns, not significant.

Article Snippet: Plates were then loaded with mouse anti-human Fc specific IgG1, IgG2, IgG3 and IgG4 that were conjugated with HRP (Southern Biotech) and incubated for another hour at room temperature (RT).

Techniques: Comparison, Infection

Journal: Frontiers in Immunology

Article Title: Spike specific IgG3 and nucleocapsid IgG response in serum serve as distinguishing immunological markers between SARS-CoV-2 infection and vaccination

doi: 10.3389/fimmu.2025.1518915

Figure Lengend Snippet: RBD specific IgG and its subclasses after COVID-19 vaccination. RBD specific IgG (A) and RBD specific IgG1 (B) , IgG2 (C) , IgG3 (D) , IgG4 (E) was analyzed in serum samples taken from COVID-19 vaccines (n=42). Samples were collected before vaccination (Pre), 1 month (post-dose 1), 2/3 Months (post-dose 2) and 6 months (post-6 M) after the first vaccination. Each symbol represents one participant where brown dots indicate mRNA and blue dots indicate Covishield, vaccine recipients. Statistical analysis was performed between pre and post-vaccination using the paired t test. **P <0.001, ***P <0.0001, ns, not significant.

Article Snippet: Plates were then loaded with mouse anti-human Fc specific IgG1, IgG2, IgG3 and IgG4 that were conjugated with HRP (Southern Biotech) and incubated for another hour at room temperature (RT).

Techniques: Vaccines

Journal: Frontiers in Immunology

Article Title: Spike specific IgG3 and nucleocapsid IgG response in serum serve as distinguishing immunological markers between SARS-CoV-2 infection and vaccination

doi: 10.3389/fimmu.2025.1518915

Figure Lengend Snippet: Boosting up of IgG1, not IgG3 after vaccination of naturally infected COVID-19 patients. RBD-specific IgG (A) , IgG1 (B) and IgG3 (C) were analyzed in serum samples from COVID-19 patients who received COVID-19 vaccines after 90 days post-infection (n=39). Samples were collected after infection and vaccination at different time interval are indicated below the graph. Each symbol represents one individual. The black dots indicate the time points before vaccination and the red dots indicates the time points after administration of vaccine. Statistical analyses were performed between prior vaccination (Day-28) and after vaccination using the Man-Whitney test. *P <0.05, **P <0.001, ***P <0.0001, ns, not significant.

Article Snippet: Plates were then loaded with mouse anti-human Fc specific IgG1, IgG2, IgG3 and IgG4 that were conjugated with HRP (Southern Biotech) and incubated for another hour at room temperature (RT).

Techniques: Infection, Vaccines

Journal: Frontiers in Immunology

Article Title: Spike specific IgG3 and nucleocapsid IgG response in serum serve as distinguishing immunological markers between SARS-CoV-2 infection and vaccination

doi: 10.3389/fimmu.2025.1518915

Figure Lengend Snippet: Boosting up of both IgG1 and IgG3 in breakthrough infection after primary vaccination. RBD-specific IgG (A) , IgG1 (B) and IgG3 (C) were analyzed in serum samples before (red dots) and after (black) breakthrough infection of COVID-19 vaccinated individuals (n=14). Each symbol represents one participant. Statistical analysis was performed between before and after breakthrough infection by using the Wilcoxon test. *P <0.05, **P <0.001, ***P <0.001.

Article Snippet: Plates were then loaded with mouse anti-human Fc specific IgG1, IgG2, IgG3 and IgG4 that were conjugated with HRP (Southern Biotech) and incubated for another hour at room temperature (RT).

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Spike specific IgG3 and nucleocapsid IgG response in serum serve as distinguishing immunological markers between SARS-CoV-2 infection and vaccination

doi: 10.3389/fimmu.2025.1518915

Figure Lengend Snippet: Nucleocapsid specific IgG responses after natural infection and vaccination. Nucleocapsid specific IgG antibody was measured in naturally infected COVID-19 patients (n=39) and healthy controls (n=20) (A) and COVID-19 vaccinated individuals (n=28) (B) . Samples were analyzed at day-1,7 and 28 after infection are indicated below graph (A) For vaccinated individuals, samples were analyzed before vaccination (pre) and after 1 month (post-dose 1) and 2/3 Months (post-dose 2) of vaccination indicated below graph (B) Each symbol represents one participant. In (A) , each dot represents one patient: asymptomatic (black), mild (green), moderate (blue) and severe (orange). In (B) , brown dots indicate mRNA and blue dots indicate Covishield, vaccine recipients. Statistical analysis was performed between healthy controls and infected patients using the Man-Whitney test and between pre and post vaccinated individuals using Wilcoxon test. **P <0.001, ***P <0.0001, ns, not significant..

Article Snippet: Plates were then loaded with mouse anti-human Fc specific IgG1, IgG2, IgG3 and IgG4 that were conjugated with HRP (Southern Biotech) and incubated for another hour at room temperature (RT).

Techniques: Infection

Journal: medRxiv

Article Title: Cross-binding antibodies capable of neutralizing diverse orthohantaviruses are produced in response to Puumala virus infection

doi: 10.1101/2025.03.04.25323331

Figure Lengend Snippet: Following an incubation period of 1-6 weeks, HFRS symptoms manifest as five distinct phases: febrile, hypotensive, oliguric, polyuric, and convalescent. The average time for which each stage lasts is shown, in addition to the symptoms associated with each phase. Serum creatinine, a measure of kidney injury, is displayed, as is urine output as a marker of kidney function. IgM and IgG titers are also shown. The four time points at which patient sera were sampled are also shown in relation to each disease phase. Time point 1 (T1) was sampled six days post-symptom onset (PSO), range: 2-11 days PSO). Time point 2 (T2) was sampled 12 days PSO (range: 8-20 days PSO). Time point 3 (T3) was sampled 28 PSO (range: 21-64 days PSO). Time point 4 (T4) was sampled 197 days PSO (range: 180-256 days PSO). Adapted from Avsic-Zupanc et al. . Created in BioRender. Clark, J. (2025) https://BioRender.com/p36u208

Article Snippet: When assaying other immunoglobulin subtypes, anti-human IgA HRP (Sigma-Aldrich, A0295, 1:3000), anti-human IgM HRP (SouthernBiotech, 2020-05, 1:3000), anti-human IgG1 (Southern Biotech, 9054-05, 1:5000), anti-human IgG2 (Southern Biotech, 9060-05,1:5000), anti-human IgG3 (Southern Biotech, 9210-0, 1:5000), or anti-human IgG4 (Southern Biotech, 9200-05, 1:5000) were used.

Techniques: Incubation, Marker

Journal: medRxiv

Article Title: Cross-binding antibodies capable of neutralizing diverse orthohantaviruses are produced in response to Puumala virus infection

doi: 10.1101/2025.03.04.25323331

Figure Lengend Snippet: ELISAs were carried out using patient sera and plates coated with ultracentrifuge purified rVSV expressing the GnGc of A) PUUV, B) SEOV, C) DOBV, D) HTNV, E) SNV, or F) ANDV. Wild-type VSV G) was used as a negative control. IgG titers are shown as area under the curve (AUC) with the geometric mean and geometric standard deviation denoted by error bars. 11 age and sex matched healthy control sera were included as negative controls and an in-house, anti-PUUV monoclonal antibody was used as a positive control. AUC values were log10 transformed and statistical significance was interrogated via ordinary one-way ANOVA using a Tukey’s multiple comparisons test. Only comparisons with p ≤ 0.05 are shown. Limits of detection (LOD) were calculated for each antigen using the average + 3x standard deviations of the AUC of the negative control sera and are shown as dotted lines on each graph. The percentage of serum samples with LODs above the limit of detection are shown as pie charts, with the percentage of positive samples displayed in green. The n for each time point is displayed below each pie chart.

Article Snippet: When assaying other immunoglobulin subtypes, anti-human IgA HRP (Sigma-Aldrich, A0295, 1:3000), anti-human IgM HRP (SouthernBiotech, 2020-05, 1:3000), anti-human IgG1 (Southern Biotech, 9054-05, 1:5000), anti-human IgG2 (Southern Biotech, 9060-05,1:5000), anti-human IgG3 (Southern Biotech, 9210-0, 1:5000), or anti-human IgG4 (Southern Biotech, 9200-05, 1:5000) were used.

Techniques: Purification, Expressing, Negative Control, Standard Deviation, Control, Positive Control, Transformation Assay

Journal: medRxiv

Article Title: Cross-binding antibodies capable of neutralizing diverse orthohantaviruses are produced in response to Puumala virus infection

doi: 10.1101/2025.03.04.25323331

Figure Lengend Snippet: ELISAs were carried out using patient sera and plates coated with recombinant truncated, Gn of A) PUUV and B) SEOV. IgG titers are shown as area under the curve (AUC) with the geometric mean and geometric standard deviation denoted by error bars. 11 age and sex matched healthy control sera were included as negative controls and an in-house, anti-PUUV monoclonal antibody was used as a positive control. AUC values were log10 transformed and statistical significance was interrogated via ordinary one-way ANOVA using a Tukey’s multiple comparisons test. Only comparisons with p ≤ 0.05 are shown. Limits of detection (LOD) were calculated for each antigen using the average + 3x standard deviations of the AUC of the negative control sera and are shown as dotted lines on each graph. The percentage of serum samples with LODs above the limit of detection are shown as pie charts, with the percentage of positive samples displayed in green. The n for each time point is displayed below each pie chart.

Article Snippet: When assaying other immunoglobulin subtypes, anti-human IgA HRP (Sigma-Aldrich, A0295, 1:3000), anti-human IgM HRP (SouthernBiotech, 2020-05, 1:3000), anti-human IgG1 (Southern Biotech, 9054-05, 1:5000), anti-human IgG2 (Southern Biotech, 9060-05,1:5000), anti-human IgG3 (Southern Biotech, 9210-0, 1:5000), or anti-human IgG4 (Southern Biotech, 9200-05, 1:5000) were used.

Techniques: Recombinant, Standard Deviation, Control, Positive Control, Transformation Assay, Negative Control

Journal: medRxiv

Article Title: Cross-binding antibodies capable of neutralizing diverse orthohantaviruses are produced in response to Puumala virus infection

doi: 10.1101/2025.03.04.25323331

Figure Lengend Snippet: ELISAs were carried out using patient sera to investigate A) anti-PUUV IgM, B) anti-PUUV IgA, C) anti-SEOV IgM, and D) anti-SEOV IgA. IgG subtype ELISAs were performed to investigate the quantities of anti-PUUV E) IgG1, F) IgG2, G) IgG3, or H) IgG4. I) A schematic of the luciferase-based reporter assay, which was utilized to characterize the effector activity promoted by patient sera. RVSV-PUUV infected Huh.75 cells were incubated with patient sera and reporter cells that express the FcγRIIIa receptor, coupled to a gene expression pathway that results in the production of luciferase. J ) The FcγRIIIa activity of the sera re shown as AUCs, with the mean and standard deviation denoted by error bars. 11 age and sex matched healthy control sera were included as negative controls, and an in-house, anti-PUUV monoclonal antibody was used as a positive control. AUC values were log10 transformed and statistical significance was interrogated via ordinary one-way ANOVA using a Tukey’s multiple comparisons test. Only comparisons with p ≤ 0.05 are shown. Limits of detection (LOD) are shown as dotted lines on each graph. The percentage of serum samples with LODs above the limit of detection are shown as pie charts with the percentage of positive samples displayed in green. The n for each time point is displayed below each pie chart.

Article Snippet: When assaying other immunoglobulin subtypes, anti-human IgA HRP (Sigma-Aldrich, A0295, 1:3000), anti-human IgM HRP (SouthernBiotech, 2020-05, 1:3000), anti-human IgG1 (Southern Biotech, 9054-05, 1:5000), anti-human IgG2 (Southern Biotech, 9060-05,1:5000), anti-human IgG3 (Southern Biotech, 9210-0, 1:5000), or anti-human IgG4 (Southern Biotech, 9200-05, 1:5000) were used.

Techniques: Luciferase, Reporter Assay, Activity Assay, Infection, Incubation, Gene Expression, Standard Deviation, Control, Positive Control, Transformation Assay

Journal: Microbiology Spectrum

Article Title: A third COVID-19 vaccine dose in kidney transplant recipients induces antibody response to vaccine and Omicron variants but shows limited Ig subclass switching

doi: 10.1128/spectrum.02190-24

Figure Lengend Snippet: Serum IgG and nAb responses to SARS-CoV-2 ancestral and Omicron variants in KTRs in comparison to HCs. Total SARS-CoV-2 Spike (S)-specific IgG ( A ) and neutralizing antibody (nAb) ( B ) against the ancestral strain (red), Omicron BA.1 (green), and Omicron BA.5 (blue) variants measured in KTRs ( n = 81) or HCs ( n = 11) who were previously vaccinated with the initial two-dose mRNA vaccine series prior to dose 3 (pre-D3), 30 days post-dose 3 (30 days post-D3), and 90 days post-dose 3 (90 days post-D3). Total anti-S IgG ( C ) and nAb levels ( D ) at 30 days post-D3 and nAb levels at 90 days post-D3 ( E ) were compared between the ancestral strain, Omicron BA.1, and Omicron BA.5 variants. Total anti-S IgG ( F ) and nAb ( G ) levels against ancestral strain, Omicron BA.1, and Omicron BA.5 variants were compared between KTR and HCs at 30 days post-D3. Dotted lines indicate the limit of detection (LOD), which is −1.52 for the anti-S IgG ELISA assay ( A, C, F ) and 0.17 for the nAb assay ( B, D, E, G ). Open circles represent non-responders with negative serological responses that fall below the LOD value. Solid triangles represent patients with a confirmed SARS-CoV-2 infection during the course of the study. The mean ± 95% CI are shown in each panel. Significance is tested using one-way repeated measures ANOVA ( A–E ), and unpaired t-tests ( F, G ). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Fold changes (X) are labeled below the significance lines. The number of positive samples out of the total number of samples tested is indicated in parentheses.

Article Snippet: For characterization of subclass-specific IgG, we used secondary IgG1 antibody (catalog 9054-05, SouthernBiotech) at a 1:4,000 dilution, IgG2 antibody (catalog 9060-05, SouthernBiotech) at a 1:4,000 dilution, IgG3 antibody (catalog 9210-05, SouthernBiotech) at a 1:4,000 dilution, and IgG4 antibody (catalog 9200-05, SouthernBiotech) at a 1:8,000 dilution.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Infection, Labeling

Journal: Microbiology Spectrum

Article Title: A third COVID-19 vaccine dose in kidney transplant recipients induces antibody response to vaccine and Omicron variants but shows limited Ig subclass switching

doi: 10.1128/spectrum.02190-24

Figure Lengend Snippet: Serum IgG subclass profile to SARS-CoV-2 ancestral strain in KTRs. SARS-CoV-2 Spike (S)-specific IgG subclasses (IgG1-4): IgG1 (yellow), IgG2 (blue), IgG3 (green), and IgG4 (purple) against ancestral strain were measured in KTRs ( n = 81) prior to dose 3 (pre-D3), 30 days post-dose 3 (30 days post-D3), and 90 days post-dose 3 (90 days post-D3) ( A ). Anti-S IgG subclasses antibody levels against SARS-CoV-2 ancestral strain were compared at 30 days post-D3 ( B ) and 90 days post-D3 ( C ) in KTRs with anti-S total IgG (red) as a reference on the left of both panels. A summary panel of anti-S IgG subclass-specific antibody levels against ancestral strain is shown and connected by lines to show the changes of serological responses in IgG subclasses from pre-D3, 30 days post-D3, to 90 days post-D3 ( D ). Comparison of anti-S IgG subclass-specific antibody levels against ancestral strain was made between KTRs and healthy controls (HCs) ( n = 11) at 30 days post-D3 ( E ). Dotted lines indicate the limit of detection (LOD), which is −3.00 for the subclass-specific IgG ELISA assay. Open circles represent non-responders with negative serological responses that fall below the LOD value. olid triangles represent patients with a confirmed SARS-CoV-2 infection during the course of the study. The mean ± 95% CI are shown in each panel. Significance is tested using mixed-effects model ( A ), one-way repeated measures ANOVA ( B, C ), and unpaired t -test ( E ). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. Fold changes (X) are labeled below the significance lines. The number of positive samples out of the total number of samples tested is indicated in parentheses.

Article Snippet: For characterization of subclass-specific IgG, we used secondary IgG1 antibody (catalog 9054-05, SouthernBiotech) at a 1:4,000 dilution, IgG2 antibody (catalog 9060-05, SouthernBiotech) at a 1:4,000 dilution, IgG3 antibody (catalog 9210-05, SouthernBiotech) at a 1:4,000 dilution, and IgG4 antibody (catalog 9200-05, SouthernBiotech) at a 1:8,000 dilution.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Infection, Labeling