fcγriia h167  (R&D Systems)

Journal: Antibody Therapeutics

Article Title: The influence of variable-heavy chain families on IgG 2 , 3 , 4 , FcγRs and B-cell superantigens protein G and L binding using biolayer interferometry

doi: 10.1093/abt/tbad016

Figure Lengend Snippet: BLI measurement of pertumuzab IgG 3 V H 1–V H 7 variants binding immobilized FcγRs (FcγRIa, FcγRIIa H167 , FcγRIIa R167 , FcγRIIb/c, FcγRIIIa F176 , FcγRIIIa V176 , FcγRIIIb NA1 and FcγRIIIb NA2 ). K D (nM), ka (×10 4 (1/Ms)) and kd (×10 −4 (1/s)) are all presented; PR indicates a poor response (RU below 0.3); all experiments were performed in triplicates with the standard errors shown

Article Snippet: Commercial Avi-tagged receptors FcγRIa (Sino Biological Inc. Cat: 10256-H27H-B), FcγRIIa H167 (Acro Biosystems Cat: CDA-H82E6 FcγRIIa R167 (Acro Biosystems Cat: CDA-H82E7), FcγRIIb/c (Acro Biosystems Cat: CDB-H82E0), FcγRIIIa F176 (Acro Biosystems Cat: CDA-H82E8), FcγRIIIa V176 Acro Biosystems Cat: CDA-H82E9), FcγRIIIb NA1 (Acro Biosystems Cat: CDB-H82E4) and FcγRIIIb NA2 (Sino Biological Inc. Cat: 11046-H27H-B) were immobilized onto streptavidin biosensors (Satorius Cat: 18-5020) at 200 nM and subjected to free-floating IgG subtype variants (concentration dilution: 100, 50, 25, 12.5 and 6.25 nM) to obtain the rate of association (ka: × 10 4 (1/Ms)), dissociation (kd: × 10 −4 (1/s)) and equilibrium dissociation constant (K D : nM).

Techniques: Binding Assay

Journal: Antibody Therapeutics

Article Title: The influence of variable-heavy chain families on IgG 2 , 3 , 4 , FcγRs and B-cell superantigens protein G and L binding using biolayer interferometry

doi: 10.1093/abt/tbad016

Figure Lengend Snippet: BLI measurement of pertumuzab IgG 2 V H 1–V H 7 variants binding immobilized FcγRs (FcγRIa, FcγRIIa H167 , FcγRIIa R167 , FcγRIIb/c, FcγRIIIa F176 , FcγRIIIa V176 , FcγRIIIb NA1 and FcγRIIIb NA2 ). K D (nM), ka (×10 4 (1/Ms)) and kd (×10 −4 (1/s)) are all presented; PR indicates a poor response (RU below 0.3); all experiments were performed in triplicates with the standard errors shown

Article Snippet: Commercial Avi-tagged receptors FcγRIa (Sino Biological Inc. Cat: 10256-H27H-B), FcγRIIa H167 (Acro Biosystems Cat: CDA-H82E6 FcγRIIa R167 (Acro Biosystems Cat: CDA-H82E7), FcγRIIb/c (Acro Biosystems Cat: CDB-H82E0), FcγRIIIa F176 (Acro Biosystems Cat: CDA-H82E8), FcγRIIIa V176 Acro Biosystems Cat: CDA-H82E9), FcγRIIIb NA1 (Acro Biosystems Cat: CDB-H82E4) and FcγRIIIb NA2 (Sino Biological Inc. Cat: 11046-H27H-B) were immobilized onto streptavidin biosensors (Satorius Cat: 18-5020) at 200 nM and subjected to free-floating IgG subtype variants (concentration dilution: 100, 50, 25, 12.5 and 6.25 nM) to obtain the rate of association (ka: × 10 4 (1/Ms)), dissociation (kd: × 10 −4 (1/s)) and equilibrium dissociation constant (K D : nM).

Techniques: Binding Assay

Journal: Antibody Therapeutics

Article Title: The influence of variable-heavy chain families on IgG 2 , 3 , 4 , FcγRs and B-cell superantigens protein G and L binding using biolayer interferometry

doi: 10.1093/abt/tbad016

Figure Lengend Snippet: BLI measurement of pertumuzab IgG 4 V H 1–V H 7 variants binding immobilized FcγRs (FcγRIa, FcγRIIa H167 , FcγRIIa R167 , FcγRIIb/c, FcγRIIIa F176 , FcγRIIIa V176 , FcγRIIIb NA1 and FcγRIIIb NA2 ); K D (nM), ka (×10 4 (1/Ms)) and kd (×10 −4 (1/s)) are all presented; PR indicates a poor response (RU below 0.3); all experiments were performed in triplicates with the standard errors shown

Article Snippet: Commercial Avi-tagged receptors FcγRIa (Sino Biological Inc. Cat: 10256-H27H-B), FcγRIIa H167 (Acro Biosystems Cat: CDA-H82E6 FcγRIIa R167 (Acro Biosystems Cat: CDA-H82E7), FcγRIIb/c (Acro Biosystems Cat: CDB-H82E0), FcγRIIIa F176 (Acro Biosystems Cat: CDA-H82E8), FcγRIIIa V176 Acro Biosystems Cat: CDA-H82E9), FcγRIIIb NA1 (Acro Biosystems Cat: CDB-H82E4) and FcγRIIIb NA2 (Sino Biological Inc. Cat: 11046-H27H-B) were immobilized onto streptavidin biosensors (Satorius Cat: 18-5020) at 200 nM and subjected to free-floating IgG subtype variants (concentration dilution: 100, 50, 25, 12.5 and 6.25 nM) to obtain the rate of association (ka: × 10 4 (1/Ms)), dissociation (kd: × 10 −4 (1/s)) and equilibrium dissociation constant (K D : nM).

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Diverging maternal and infant cord antibody functions from SARS-CoV-2 infection and vaccination in pregnancy

doi: 10.1101/2023.05.01.538955

Figure Lengend Snippet: A subset of infant cord SARS-CoV-2 neutralizing and RBD Fc effector antibody functions is enhanced with vaccination compared to infection in pregnancy. The medians (bars) for cord sample ( A ) neutralization (FRNT50) against SARS-CoV-2 WA1 (infection n=14, vaccine n=13, vaccine+infection n=19), Delta and Omicron viruses (infection n=12, vaccine n=8, vaccine+infection n=14), ( B ) RBD antibody-dependent natural killer cell activation (ADNKA) by CD107a, IFNγ, and TNFα, ( C ) RBD antibody-dependent complement deposition (ADCD), ( D ) RBD antibody-dependent cellular phagocytosis (ADCP), and ( E ) relative binding of RBD-specific antibodies to FcγRIIIa/CD16a, FcγRIIa/CD32a, FcγRIIb/CD32b and FcRN are shown. For B-E, sample sizes are infection n=20, vaccine n=18, vaccine+infection n=27. P-values for A-E are adjusted for maternal age and body mass index using linear regression. ( F ) The magnitude of cord functions are summarized in the radar plot. Each line represents the median Z-scored data for each clinical group. ( G ) The proportion of detectable functions was used to categorize individuals as a high, medium or low responder. The percentages of each type of responder within each clinical group depict the polyfunctional antibody breadth.

Article Snippet: Recombinant Fc receptors (FcγRIIIa/CD16a, FcγRIIa/CD32a H167, FcγRIIb/CD32b, Neonatal Fc receptor/FcRN) (R&D Systems) were labeled with PE (Abcam) per manufacturer’s instructions, added (1μg/mL) to bead bound antigen-specific immune complexes.

Techniques: Infection, Neutralization, Activation Assay, Binding Assay

Journal: bioRxiv

Article Title: Diverging maternal and infant cord antibody functions from SARS-CoV-2 infection and vaccination in pregnancy

doi: 10.1101/2023.05.01.538955

Figure Lengend Snippet: A subset of maternal SARS-CoV-2 neutralizing and RBD Fc effector antibody functions is enhanced with vaccination compared to infection in pregnancy. The medians (bars) of the maternal pair of the cord samples in in ( A ) neutralization (FRNT50) against SARS-CoV-2 WA1 (infection n=14, vaccine n=13, vaccine+infection n=19), Delta and Omicron viruses (infection n=12, vaccine n=8, vaccine+infection n=14), ( B ) RBD antibody-dependent natural killer cell activation (ADNKA) as measured by CD107a, IFNγ, and TNFα, ( C ) RBD antibody-dependent complement deposition (ADCD), ( D ) RBD antibody-dependent cellular phagocytosis (ADCP), and ( E ) relative binding of RBD-specific antibodies to FcγRIIIa/CD16a, FcγRIIa/CD32a, FcγRIIb/CD32b and FcRN. For B-E, sample sizes are infection n=22, vaccine n=19, vaccine+infection n=28. P-values for A-E are adjusted for maternal age and body mass index using linear regression. ( F ) The magnitude of maternal functions are summarized in the radar plot. Each line represents the median Z-scored data for each clinical group. ( G ) The proportion of detectable functions was used to categorize individuals as a high, medium or low responder. The percentages of each type of responder within each clinical group depict the polyfunctional antibody breadth.

Article Snippet: Recombinant Fc receptors (FcγRIIIa/CD16a, FcγRIIa/CD32a H167, FcγRIIb/CD32b, Neonatal Fc receptor/FcRN) (R&D Systems) were labeled with PE (Abcam) per manufacturer’s instructions, added (1μg/mL) to bead bound antigen-specific immune complexes.

Techniques: Infection, Neutralization, Activation Assay, Binding Assay

Journal: bioRxiv

Article Title: Diverging maternal and infant cord antibody functions from SARS-CoV-2 infection and vaccination in pregnancy

doi: 10.1101/2023.05.01.538955

Figure Lengend Snippet: SARS-CoV-2 neutralizing and RBD Fc effector functions are differentially transferred across the placenta. ( A ) Neutralization against live SARS-CoV-2 WA1, variant Delta and Omicron, ( B ) RBD ADNKA, ( C ) RBD ADCD, ( D ) RBD ADCP, and ( E ) relative binding of RBD-specific IgG to FcγRIIIa/CD16a, FcγRIIa/CD32a, FcγRIIb/CD32b and FcRN are compared with the values of the medians for maternal (grey) and matched cord (blue) samples listed below. Statistical significance was calculated by Wilcoxon-matched pairs test. ( F ) Antibody function transfer ratios (the proportion of cord to maternal levels) are shown with medians (bars), interquartile ranges (boxes), and ranges (whiskers).

Article Snippet: Recombinant Fc receptors (FcγRIIIa/CD16a, FcγRIIa/CD32a H167, FcγRIIb/CD32b, Neonatal Fc receptor/FcRN) (R&D Systems) were labeled with PE (Abcam) per manufacturer’s instructions, added (1μg/mL) to bead bound antigen-specific immune complexes.

Techniques: Neutralization, Variant Assay, Binding Assay

Journal: bioRxiv

Article Title: Diverging maternal and infant cord antibody functions from SARS-CoV-2 infection and vaccination in pregnancy

doi: 10.1101/2023.05.01.538955

Figure Lengend Snippet: Antibody functions highlight the effect of differential immune exposure in pregnancy while glycosylation marks diverging maternal and infant cord responses. Bubble plots ( A ) show the correlation between neutralizing activities against SARS-CoV-2 WA1, Delta and Omicron and RBD-specific Fc effector functions of antibody-dependent natural killer cell activation (CD107a, IFNγ, TNFα), antibody-dependent complement deposition (ADCD), antibody-dependent cellular phagocytosis (ADCP) and relative binding to FcγR (FcγRIIIa/CD16a, FcγRIIa/CD32a, FcγRIIb/CD32b, FcRN). The Spearman’s rank correlation coefficient is shown by color and significance (-log p) by size with those p<0.05 depicted. Principle-component analysis (PCA) using 38 SARS-CoV-2 antibody functions and features show separations between infection and vaccine clinical groups and maternal and cord responses. Each symbol in the ( B ) score plot represents a single maternal or cord sample. Each antibody feature is represented in the ( C ) loadings plot, where its location reflects the distribution of the individual samples in the ( B ) score plot.

Article Snippet: Recombinant Fc receptors (FcγRIIIa/CD16a, FcγRIIa/CD32a H167, FcγRIIb/CD32b, Neonatal Fc receptor/FcRN) (R&D Systems) were labeled with PE (Abcam) per manufacturer’s instructions, added (1μg/mL) to bead bound antigen-specific immune complexes.

Techniques: Activation Assay, Binding Assay, Infection