Journal: mAbs

Article Title: Systematic analysis of Fc mutations designed to reduce binding to Fc-gamma receptors

doi: 10.1080/19420862.2024.2402701

Figure Lengend Snippet:

Article Snippet: FcγRIIa (CD32a) 131 R allele , human , Sino Biological , 10374-H08H.

Techniques:

Journal: mAbs

Article Title: Fc-mediated activity of EGFR x c-Met bispecific antibody JNJ-61186372 enhanced killing of lung cancer cells

doi: 10.1080/19420862.2016.1249079

Figure Lengend Snippet: Increased FcγRIIIa binding of low-and hemi-fucosylated forms of JNJ-61186372. FcγRI (A), FcγRIIa (B), and FcγRIIIa (C) binding by antibodies produced in normal fucose (NF) or low fucose (LF) cell line was assessed by competitive Alpha Screen and compared to a wild type IgG1 control antibody (closed circle). JNJ-61186372 – NF (closed square), JNJ-61186372 – LF (closed up triangle), EGFR x inert arm –LF (closed down triangle), c-Met x inert arm – LF (open up triangle), EGFR (LF) x c-Met (NF) (open down triangle), EGFR (NF) x c-Met (LF) (open diamond), JNJ-61186372 – IgG2σ (open circle). Representative data from 3 replicate experiments is shown.

Article Snippet: His-tagged FcγRI (R&D Systems, cat no. 1257-FC), FcγRIIa (R&D Systems, cat no. 1330-CD/CF) or FcγRIIIa (R&D Systems, cat no. 4325-FC) were captured on nickel chelate acceptor beads (PerkinElmer, cat. no. CUSM0220400EA).

Techniques: Binding Assay, Produced, Control

Journal: mAbs

Article Title: Fc-mediated activity of EGFR x c-Met bispecific antibody JNJ-61186372 enhanced killing of lung cancer cells

doi: 10.1080/19420862.2016.1249079

Figure Lengend Snippet: Concentration of antibody (nM) to elicit 50% of maximal FcγR binding activity (IC 50 ). Data presented in mean ± SEM of 2–3 independent experiments. *Difference between JNJ-61186372 - LF and JNJ-61186372 – NF is statistically significant (p value = 0.0001).

Article Snippet: His-tagged FcγRI (R&D Systems, cat no. 1257-FC), FcγRIIa (R&D Systems, cat no. 1330-CD/CF) or FcγRIIIa (R&D Systems, cat no. 4325-FC) were captured on nickel chelate acceptor beads (PerkinElmer, cat. no. CUSM0220400EA).

Techniques: Concentration Assay, Binding Assay, Activity Assay, Significance Assay

Journal: mAbs

Article Title: Fc-mediated activity of EGFR x c-Met bispecific antibody JNJ-61186372 enhanced killing of lung cancer cells

doi: 10.1080/19420862.2016.1249079

Figure Lengend Snippet: Fold change to multiple FcγR binding (IC 50 ) of different antibodies to that of JNJ-61186372 – NF.

Article Snippet: His-tagged FcγRI (R&D Systems, cat no. 1257-FC), FcγRIIa (R&D Systems, cat no. 1330-CD/CF) or FcγRIIIa (R&D Systems, cat no. 4325-FC) were captured on nickel chelate acceptor beads (PerkinElmer, cat. no. CUSM0220400EA).

Techniques: Binding Assay

Journal: Cell Reports

Article Title: Antibodies targeting conserved non-canonical antigens and endemic coronaviruses associate with favorable outcomes in severe COVID-19

doi: 10.1016/j.celrep.2022.111020

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Article Snippet: CD32a , Acrobiosystems , Cat #CDA-H82E6-25ug.

Techniques: Recombinant, Plasmid Preparation, Software

Journal: Cancer Biology & Therapy

Article Title: CA125 suppresses amatuximab immune-effector function and elevated serum levels are associated with reduced clinical response in first line mesothelioma patients

doi: 10.1080/15384047.2018.1449614

Figure Lengend Snippet: Cell-based and ELISA-based assays to test for CA125 binding to amatuximab and effect on antibody- CD16a Fc-γ receptor engagement. A) Biotinylated amatuximab and its (Fab’) 2 fragment, but not its Fc fragment bind immobilized CA125. 96-well plates were coated with 15 KU/mL CA125 or human serum albumin (control) and probed with biotin-labeled amatuxiumab, (Fab’) 2 or Fc fragments. *** p < 0.00002; ***** p < 0.000002. B) Testing the effect of CA125 on Jurkat-CD16a binding using BRA assay. Jurkat-CD16a cells were incubated in wells coated with amatuximab and incubated with or without sCA125. CA125 inhibited Jurkat-CD16a-amatuximab binding (top row). Farletuzumab (middle row) was used as a positive control and humanized anti-tissue factor (TF) antibody (bottom row) was used as a negative control for the assay. Shown are duplicate experiments for each condition. C) CA125 suppresses Fc receptor binding to amatuximab. Amatuximab was incubated alone or with sCA125 and probed with biotinylated-CD16a, -CD32a or -CD64a Fc-γ receptors. As shown, CA125 caused a significant decrease of CD16a binding to amatuximab (50%, p = 0.0000009). Reduction in amatuximab binding to CD32a Fc receptors was also significant (29%, p < 0.003) while no inhibition was observed by CA125 on amatuximab binding to CD64a Fc receptor (0%, p = 0.380). Similar reactions were probed with anti-human IgG-HRP to ensure incubation of amatuximab with CA125 did not result in less amatuximab binding to wells (last set of bars). D) FcRn which also binds to the Fc domain of amatuximab was biotinylated and used as probe to determine if CA125 interfered with its ability to bind amatuximab Fc domain. As shown, FcRn binding is not affected by CA125 binding to amatuximab.

Article Snippet: Recombinant human CD16a-, CD32a- and CD64a-biotin probes were purchased from Sino Biological Inc. (Beijing, China).

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Labeling, Incubation, Positive Control, Negative Control, Inhibition