igg3 reference antibodies  (Athens Research)

Journal: Vaccines

Article Title: FLU-v, a Broad-Spectrum Peptide-Based Influenza Vaccine, Induces NK Cell Activating IgG1 and IgG3 Subclass Antibodies in Humans

doi: 10.3390/vaccines13111084

Figure Lengend Snippet: FLU-v IgG1 ( A ) and IgG3 ( B ) concentrations across all groups on days 0, 42, and 180. Data from the following groups are shown: adjuvanted FLU-v ( n = 52), non-adjuvanted FLU-v ( n = 58), adjuvanted placebo ( n = 26), and non-adjuvanted placebo ( n = 32). Box and whisker plot showing all individual values, 25th and 75th percentiles, and median. Whiskers represent ranges. The Mann–Whitney U test was used to compare different groups at each time point. The Wilcoxon signed-rank sum test was used to compare differences between time points within each group. Only significant differences are shown.

Article Snippet: Wells in the standard rows were coated with human IgG1 or IgG3 reference antibodies (16-16-090707-1/16-16-090707-3, Athens Research, Athens, GA, USA), performing double decreasing serial dilutions starting at 1000 ng/mL up to 1.95 ng/mL.

Techniques: Whisker Assay, MANN-WHITNEY

Journal: Vaccines

Article Title: FLU-v, a Broad-Spectrum Peptide-Based Influenza Vaccine, Induces NK Cell Activating IgG1 and IgG3 Subclass Antibodies in Humans

doi: 10.3390/vaccines13111084

Figure Lengend Snippet: Fold increase in FLU-v IgG1 ( A ) and IgG3 ( B ) concentrations across all groups from day 0 to days 42 and 180. Data from the following groups are shown: adjuvanted FLU-v ( n = 52), non-adjuvanted FLU-v ( n = 58), adjuvanted placebo (n = 26), and non-adjuvanted placebo ( n = 32). Box and whiskers plot showing all individual values, 25th and 75th percentiles, and median. Whiskers represent ranges. Fold change in IgG1 concentrations on day 42 and 180 is defined as the ratio of day 42 to day 0 and the ratio of day 180 to day 0, respectively. The Mann–Whitney U test was used to compare different groups at each time point. The Wilcoxon signed-rank sum test was used to compare differences between day 42 and 180 within each group. Significant differences are shown.

Article Snippet: Wells in the standard rows were coated with human IgG1 or IgG3 reference antibodies (16-16-090707-1/16-16-090707-3, Athens Research, Athens, GA, USA), performing double decreasing serial dilutions starting at 1000 ng/mL up to 1.95 ng/mL.

Techniques: MANN-WHITNEY

Journal: Vaccines

Article Title: FLU-v, a Broad-Spectrum Peptide-Based Influenza Vaccine, Induces NK Cell Activating IgG1 and IgG3 Subclass Antibodies in Humans

doi: 10.3390/vaccines13111084

Figure Lengend Snippet: Antibody-mediated NK cell activation after adjuvanted and non-adjuvanted FLU-v vaccination. For this analysis, a subset of paired samples ( n = 50) from day 0 and day 42 was selected based on high IgG1 and IgG3 fold increases. Data from the following groups are shown: adjuvanted FLU-v ( n = 32), non-adjuvanted FLU-v ( n = 16), and non-adjuvanted placebo ( n = 2). Antibody-dependent NK cell activation, or percentage of GFP + CD107a + CD16 (V176) NK-92 cells by flow cytometry, was measured in response to antibodies immobilized by plate-bound FLU-v ( A ) or non-related peptides used as negative controls ( B ). Medians ± 95% confidences intervals are shown. The Wilcoxon signed-rank sum test was used to compare differences between time points within each vaccine group. A Bonferroni test was used to correct for multiple comparisons, with a p value < 0.025 considered significant.

Article Snippet: Wells in the standard rows were coated with human IgG1 or IgG3 reference antibodies (16-16-090707-1/16-16-090707-3, Athens Research, Athens, GA, USA), performing double decreasing serial dilutions starting at 1000 ng/mL up to 1.95 ng/mL.

Techniques: Activation Assay, Flow Cytometry

Journal: Vaccines

Article Title: FLU-v, a Broad-Spectrum Peptide-Based Influenza Vaccine, Induces NK Cell Activating IgG1 and IgG3 Subclass Antibodies in Humans

doi: 10.3390/vaccines13111084

Figure Lengend Snippet: Correlation between antibody-mediated NK cell activation and FLU-v-specific IgG ( A ), IgG1 ( B ) and IgG3 ( C ) on days 0 and 42. Spearman rank order correlations were performed between the antibody-dependent NK cell activation assay, measuring the percentage of GFP + CD107a + CD16 (V176) NK-92 cells by flow cytometry, and total IgG, IgG1, and IgG3 with correlation coefficients (r) and p -values shown. IgG data for this analysis were obtained from a previous report .

Article Snippet: Wells in the standard rows were coated with human IgG1 or IgG3 reference antibodies (16-16-090707-1/16-16-090707-3, Athens Research, Athens, GA, USA), performing double decreasing serial dilutions starting at 1000 ng/mL up to 1.95 ng/mL.

Techniques: Activation Assay, Flow Cytometry

Journal: Hematology (Amsterdam, Netherlands)

Article Title: Metabolomics-based study on the significance of differential metabolite binding IgG isoforms in Hemolytic disease of newborn.

doi: 10.1080/16078454.2024.2360339

Figure Lengend Snippet: Figure 2. Abnormal levels of IgG subtypes in HDN. A. ELISA was used to measure the levels of IgG1 in the serum of newborns in each group. B. ELISA was used to measure the levels of IgG2a in the serum of newborns in each group. C. Levels of IgG2b in each group. D. Levels of IgG3 in the serum of newborns in the Healthy, HDN, and BLI groups. E. Levels of IgG4 in the serum of newborns in the three groups. * indicates a significant difference compared to the Healthy group, with statistical significance set at P < 0.05. # indicates a significant difference compared to the HDN group, with statistical significance set at P < 0.05.

Article Snippet: ELISA kits for IgG1 (E-EL-H6087, Elabscience, WuHan), IgG2a (YJ038048, YuanJu, Shanghai), IgG2b (YJ038047, YuanJu, Shanghai), IgG3 (E-EL-H6088, Elabscience, WuHan), and IgG4 (E-EL-H6186, Elabscience, WuHan) were purchased.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: A homogeneous bioluminescent immunoassay for parallel characterization of binding between a panel of antibodies and a family of Fcγ receptors

doi: 10.1038/s41598-022-15887-z

Figure Lengend Snippet: List of antibodies used in this work.

Article Snippet: 3 , Human IgG3 , Athens Research & Technology , 16-16-090707-3.

Techniques:

Journal: Scientific Reports

Article Title: A homogeneous bioluminescent immunoassay for parallel characterization of binding between a panel of antibodies and a family of Fcγ receptors

doi: 10.1038/s41598-022-15887-z

Figure Lengend Snippet: Representative data for the tracer-target titration shown for FcγRIIIa (V158). ( A ) Concentration dependent increase in luminescent signal as function of FcγRIIIa (V158)-SmBiT target and IgG-LgBiT tracer. Presence of excess human IgG competes with the tracer resulting in decrease in luminescent signal. Luminescent signal in the presence of excess human IgG is considered background and arises from random interaction between SmBiT and LgBiT. Data was generated in a 96-well plate in single replicate. ( B ) Background subtracted luminescence signal represent specific interaction between target and the tracer. ( C ) Signal/background (S/B) ratio is calculated at each tracer target concentration and a specific concentration of tracer and target is selected (shown in the Table) for subsequent assays that satisfies the Cheng-Prusoff equation and offers high luminescence signal so that assay can be performed on any luminometer.

Article Snippet: 3 , Human IgG3 , Athens Research & Technology , 16-16-090707-3.

Techniques: Titration, Concentration Assay, Generated

Journal: Scientific Reports

Article Title: A homogeneous bioluminescent immunoassay for parallel characterization of binding between a panel of antibodies and a family of Fcγ receptors

doi: 10.1038/s41598-022-15887-z

Figure Lengend Snippet: Dose dependent inhibition curves generated with six different FcγR assays. Four different set of samples were tested to show the specificity and subclass specific binding. Analytes tested are (1) human IgG subclasses IgG1, IgG2, IgG3, IgG4; (2) human IgG; (3) Fc, Fab, and F(ab) 2 domains; and (4) human serum albumin (HSA). Data represent the mean ± standard error of triplicate experiments. IC 50 (nM) values calculated from the inhibition curves are shown in the Table. IC 50 values are in nM. *For FcγRIIb IC 50 values are intended only for qualitative purposes as mentioned in the text. n.d. not determined.

Article Snippet: 3 , Human IgG3 , Athens Research & Technology , 16-16-090707-3.

Techniques: Inhibition, Generated, Binding Assay

Journal: Scientific Reports

Article Title: A homogeneous bioluminescent immunoassay for parallel characterization of binding between a panel of antibodies and a family of Fcγ receptors

doi: 10.1038/s41598-022-15887-z

Figure Lengend Snippet: Dose dependent inhibition curves for recombinant anti CD20 IgG1 (CD20), afucosylated anti CD20 (CD20 fut), and deglycosylated anti CD20 (CD20 NQ) on FcγRI, FcγRIIa (H131) and FcγRIIIa (V158). Responses for two variants of FcγRII and FcγRIII were similar therefore only one variant is shown here.

Article Snippet: 3 , Human IgG3 , Athens Research & Technology , 16-16-090707-3.

Techniques: Inhibition, Recombinant, Variant Assay

Journal: Scientific Reports

Article Title: A homogeneous bioluminescent immunoassay for parallel characterization of binding between a panel of antibodies and a family of Fcγ receptors

doi: 10.1038/s41598-022-15887-z

Figure Lengend Snippet: Dose dependent inhibition curves generated with six different NanoBiT FcγR assays. Four commercial IVIg preparation were tested. Inhibition curve for IgG, generated earlier (Fig. ), is also included for comparison. Data represent the mean ± standard error of triplicate readings. IC 50 (nM) values calculated from the inhibition curves are shown in the Table. IC 50 values are in nM. *For FcγRIIb, IC 50 values are intended only for qualitative purposes as mentioned in the text.

Article Snippet: 3 , Human IgG3 , Athens Research & Technology , 16-16-090707-3.

Techniques: Inhibition, Generated, Comparison

Journal: Scientific Reports

Article Title: A homogeneous bioluminescent immunoassay for parallel characterization of binding between a panel of antibodies and a family of Fcγ receptors

doi: 10.1038/s41598-022-15887-z

Figure Lengend Snippet: Scatter plot of the IC 50 values of seven IgG1 antibodies and five different polyclonal IgG plotted two different ways. Lines are the median values. These plots demonstrate the potential advantage of matrix-screening method as a visual aide during large antibody discovery campaigns.

Article Snippet: 3 , Human IgG3 , Athens Research & Technology , 16-16-090707-3.

Techniques:

Journal: Scientific Reports

Article Title: Focused peptide library screening as a route to a superior affinity ligand for antibody purification

doi: 10.1038/s41598-021-91208-0

Figure Lengend Snippet: Binding of pIT2 and pIT2-SL min19Fc-Q6D-displaying virions to human IgG-coated and blank wells (skimmed milk-blocked only) in phage ELISA assay. Wild-type phenotype virions (with stuffer insert harboring stop codons on pIT2 and pIT2-SL background) were used as controls.

Article Snippet: Binding of monoclonal antibodies panitumumab (IgG2; Vectibix, Amgen), nivolumab (IgG4; Opdivo, Bristol-Myers Squibb), and IgG3 from human plasma (Athens Research & Technology) to the peptide A resin was assessed.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Focused peptide library screening as a route to a superior affinity ligand for antibody purification

doi: 10.1038/s41598-021-91208-0

Figure Lengend Snippet: Assessment of relative affinities for human IgG of highly enriched clones. ( a ) Phage ELISA screen conducted with 2 × 10 9 phage particles per well. ( b ) Phage ELISA assay conducted with two-fold dilutions of phage clones that gave highest signals in ( a ) Average values and standard deviations of 3 parallel experiments are presented.

Article Snippet: Binding of monoclonal antibodies panitumumab (IgG2; Vectibix, Amgen), nivolumab (IgG4; Opdivo, Bristol-Myers Squibb), and IgG3 from human plasma (Athens Research & Technology) to the peptide A resin was assessed.

Techniques: Clone Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Focused peptide library screening as a route to a superior affinity ligand for antibody purification

doi: 10.1038/s41598-021-91208-0

Figure Lengend Snippet: SDS-PAGE analysis of eluted fractions for assessment of binding specificity. A 1:4 mixture (mass ratio) of infliximab (chimeric IgG1) and BSA was loaded on the columns. E1–E4—sequential 1.5 mL elution fractions; FT—low-through; M—protein marker (ProSieve color protein marker, Lonza).

Article Snippet: Binding of monoclonal antibodies panitumumab (IgG2; Vectibix, Amgen), nivolumab (IgG4; Opdivo, Bristol-Myers Squibb), and IgG3 from human plasma (Athens Research & Technology) to the peptide A resin was assessed.

Techniques: SDS Page, Binding Assay, Marker

Journal: Scientific Reports

Article Title: Focused peptide library screening as a route to a superior affinity ligand for antibody purification

doi: 10.1038/s41598-021-91208-0

Figure Lengend Snippet: Chromatograms showing adsorption and elution of panitumumab (IgG2; a ) and nivolumab (IgG4; b ) for peptide A-based affinity column.

Article Snippet: Binding of monoclonal antibodies panitumumab (IgG2; Vectibix, Amgen), nivolumab (IgG4; Opdivo, Bristol-Myers Squibb), and IgG3 from human plasma (Athens Research & Technology) to the peptide A resin was assessed.

Techniques: Adsorption, Affinity Column