Journal: Cancers

Article Title: Affinity Affects the Functional Potency of Anti-GD2 Antibodies by Target-Mediated Drug Disposition

doi: 10.3390/cancers17152510

Figure Lengend Snippet: Reduction of anti-GD2 antibody binding to neuroblastoma cells by soluble GD2 (sGD2). NAXI and DB (0.001–1 µg/mL) were used in a competitive binding assay to GD2 high-expressing CHLA-20 tumor cells in the presence of 10, 100, and 1000 nM sGD2. An FITC-conjugated human IgG1-specific secondary antibody was used for the detection of bound anti-GD2 mAbs. ( A ) DB ( left panels) and NAXI ( right panels) show binding (gMFI GD2) in the presence and absence of sGD2 (10–1000 nM) with linear- ( upper panels) and log-y-scales ( lower panels). ( B ) Fold decrease in the binding of anti-GD2 antibodies to neuroblastoma cells (gMFI GD2) in the presence of sGD2 (NAXI: squares; DB: circles). ( C ) Direct comparison of DB (circles) and NAXI (squares) binding to GD2-expressing neuroblastoma cells incubated with 10 nM ( left panel), 100 nM ( center panel), and 1000 nM ( right panel) sGD2. Results are shown using linear- ( upper panels) and log-scales ( lower panels). Data represent the mean of gMFI ± SEM from four independent experiments. Statistical comparisons ( A ) One-way ANOVA 1000 nM (*), 100 nM ($), and 10 nM (§) vs. 0.001–1 µg/mL. $$/§§ p < 0.01, ***/$$$/§§§ p < 0.001, and ****/$$$$ p < 0.0001 versus untreated NAXI. ( B , C ) Two-sided t -tests were used for statistical analysis, with significance indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 versus NAXI.

Article Snippet: To determine the impact of soluble target antigen, we added 100 nM and 1000 nM sGD2 (Merck, Burlington, MA, USA, 345743-500UG) in addition to the ADCC setting.

Techniques: Binding Assay, Competitive Binding Assay, Expressing, Comparison, Incubation

Journal: Cancers

Article Title: Affinity Affects the Functional Potency of Anti-GD2 Antibodies by Target-Mediated Drug Disposition

doi: 10.3390/cancers17152510

Figure Lengend Snippet: Impact of soluble GD2 (sGD2) on anti-GD2 antibody-mediated cellular cytotoxicity. Spheroids were transduced to yield a stable near-infrared fluorescence for viability tracking. Tumor spheroids were treated with 75,000 PBMCs, 0.1 µg/mL dinutuximab beta (DB), and naxitamab (NAXI) (ADCC) with and without 100 and 1000 nM sGD2 for 168 h. The area under the viability curve (AUC) was calculated for each curve, and the loss of viability results in a decrease of viability values. ( A ) Graph shows the AUC (viability values) of spheroids treated with NAXI, DB, and PBMCs (ADCC) with and without 100 or 1000 nM sGD2. ( B ) Comparison of AUC (viability values) of DB- and NAXI-mediated ADCC in the presence of 100 nM and 1000 nM sGD2. Data are shown as means from at least five independent experiments (done in four replicates) ± SEM. ( A , B ) ANOVA with appropriate post hoc test: ( A ) * p < 0.05 vs. NAXI ADCC; ns = not significant. ( B ) ** p < 0.01.

Article Snippet: To determine the impact of soluble target antigen, we added 100 nM and 1000 nM sGD2 (Merck, Burlington, MA, USA, 345743-500UG) in addition to the ADCC setting.

Techniques: Fluorescence, Comparison