Journal: Frontiers in Oncology

Article Title: A trispecific antibody targeting EGFR/cMET/VEGF-A demonstrates multiple mechanisms of action to inhibit wild-type and mutant NSCLC animal models

doi: 10.3389/fonc.2025.1533059

Figure Lengend Snippet: TAVO412 bound to NSCLC cell lines and blocked binding of EGF and HGF. The binding of TAVO412 (red open circle), Amivantamab analogue (blue open circle) and null mAb (black open circle) to (A) NCI-H292; (B) HCC827; and (C) NCI-H1975 NSCLC cell lines as analyzed by flow cytometry. See Supplementary Table S2 for corresponding EC 50 , 95% CI for EC 50 , and efficacy (span in y axis). Blocking of (D) EGF and (E) HGF from binding to HCC827 cells was assessed by flow cytometry with TAVO412 (red open circle), Amivantamab analogue (blue open circle) and null mAb (black open circle). See Supplementary Table S3 for corresponding IC 50 , 95% CI for IC 50 , and efficacy (span in Y-axis). The data from three independent experiments were expressed as the mean ± SEM of duplicate treatments. The amivantamab analogue served as a positive control molecule while the null mAb served as a negative control. The abbreviations were: gMFI, geometric mean fluorescent intensity; AF647, Alexa Fluor 647 dye; AF488, Alexa Fluor 488 dye; Ab, antibody; nM, nanomolar; EGF, epidermal growth factor; HGF, hepatocyte growth factor; SEM, standard error of the mean.

Article Snippet: The cells were then incubated with AF647 goat anti-human IgG1 Fc (Jackson ImmunoResearch, 109-605-190) in the dark for 30 min at 4°C, washed three times with FACS buffer, and resuspended in FACS buffer for flow cytometry (Beckman CytoFLEX) experiments.

Techniques: Binding Assay, Flow Cytometry, Blocking Assay, Positive Control, Negative Control

Journal: Progress in neurobiology

Article Title: Astrocytic EphB3 receptors regulate d-serine-gated synaptic plasticity and memory.

doi: 10.1016/j.pneurobio.2025.102747

Figure Lengend Snippet: Fig. 1. Identification of ephrinB3-Fc doses to induce D-serine release in acute hippocampal slices. (A) Representative NMDAR-fEPSPs traces (on the left) are the average of 90 successive sweeps before (1) and after (2) clustered ephrinB3-Fc application. Results (on the bottom) are presented as means ± s.e.m. for NMDAR- fEPSP slopes, measured 60–90 min after clustered ephrinB3-Fc application, relative to 30 min of baseline recording. This dose response relationship led us to choose the dose of 0.5 µg/ml of ephrinB3-Fc. (B, C) The bar graphs represent the level of D-serine released and the ratio of D-serine /glycine measured in the extracellular medium of hippocampal slices using laser-induced fluorescence capillary electrophoresis. (B) Results are presented as means ± s.e.m. for changes in D- serine concentration induced by either clustered IgG1-Fc (0.27 µg/ml) or ephrinB3-Fc (0.5 µg/ml) condition relative to D-serine concentration obtained in clustered IgG1-Fc condition. Application of ephrinB3-Fc during 1 h induced an increase of D-serine concentration in the extracellular space. * P < 0.05. Mann Whitney test, two-tailed.

Article Snippet: Langlais et al. Progress in Neurobiology 248 (2025) 102747 under agitation with an AffiniPure goat anti-human IgG1-Fc fragment specific antibody (1:2; Jackson ImmunoResearch).

Techniques: Fluorescence, Electrophoresis, Concentration Assay, MANN-WHITNEY, Two Tailed Test

Journal: Progress in neurobiology

Article Title: Astrocytic EphB3 receptors regulate d-serine-gated synaptic plasticity and memory.

doi: 10.1016/j.pneurobio.2025.102747

Figure Lengend Snippet: Fig. 2. EphB3 receptor activation leads to an increase of synaptic NMDARs activity. (A) Representative NMDAR-fEPSPs traces are the average of 90 successive sweeps before (1) and after (2) bath applications of either clustered IgG1-Fc (0.27 µg/ml) or ephrinB3-Fc (0.5 µg/ml) and then (3) D-serine (50 µM). While IgG1-Fc had no impact, ephrinB3-Fc induced an increase of NMDARs activity through an increase of the co-agonist binding site occupancy.(B), (C) Bar graphs summarizing results presented in (A). * P < 0.05; ** P < 0.01. Mann Whitney test, two-tailed., (B) The bar graph shows the effect of clustered chimeras on synaptic NMDAR responses. Results are presented as means ± s.e.m. for NMDAR-fEPSP slopes measured after clustered chimera application (60–90 min) relative to baseline (-30–0 min). (C) Effect of D-serine on synaptic NMDAR responses in clustered chimera condition. Results are presented as means ± s.e.m. for NMDAR-fEPSP slopes measured after D-serine application (100–130 min) relative to clustered chimera condition (60–90 min). (D) Representative NMDAR-fEPSPs traces before (1) and during (2) bath applications of first D-serine and then (3) clustered ephrinB3-Fc. This occlusion experiment, represented on the plot, shows the specificity of action of ephrinB3-Fc on the release of D-serine. EphrinB3-Fc had no additional effect of the co-agonist binding site occupancy as it was first fully saturated by exogenous D- serine (50 µM). (E) Each connected circle represents paired mean NMDAR-fEPSP slope, for each individual experiment averaged in (D) measured in D-serine condition before (10–40 min) and after clustered ephrinB3-Fc application (100–130 min), relative to baseline (-30–0 min). n.s. (P > 0.05). Wilcoxon signed-rank test.

Article Snippet: Langlais et al. Progress in Neurobiology 248 (2025) 102747 under agitation with an AffiniPure goat anti-human IgG1-Fc fragment specific antibody (1:2; Jackson ImmunoResearch).

Techniques: Activation Assay, Activity Assay, Binding Assay, MANN-WHITNEY, Two Tailed Test

Journal: Progress in neurobiology

Article Title: Astrocytic EphB3 receptors regulate d-serine-gated synaptic plasticity and memory.

doi: 10.1016/j.pneurobio.2025.102747

Figure Lengend Snippet: Fig. 4. Endogenous EphB3-ephrinB3 interaction regulates synaptic D-serine availability and thus synaptic NMDAR activity. (A) Representative NMDAR-fEPSPs traces are the average of 60 successive sweeps before (1) and after (2) bath applications of unclustered IgG1-Fc (1.08 µg/ml) or ephrinB3-Fc (2.0 µg/ml) and then (3) D- serine (50 µM). The inhibition of endogenous EphB3-ephrinB3 interaction by unclustered ephrinB3-Fc induced a decrease of the activity of NMDARs. This effect was rescued by exogenous application of D-serine (50 µM).(B), (C) Bar graphs summarize results presented in (A). * P < 0.05. Mann Whitney test, two-tailed. (B) The bar graph represents the effect of unclustered chimeras on synaptic NMDAR activity. Results are presented as means ± s.e.m. for NMDAR-fEPSP slopes measured after unclustered chimera application (70–90 min) relative to baseline (-20–0 min). (C) This bar graph shows the effect of D-serine on synaptic NMDAR activity in unclustered chimera condition. Results are presented as means ± s.e.m. for NMDAR-fEPSP slopes measured after D-serine application (110–130 min) relative to unclustered chimera condition (70–90 min).(D) Representative NMDAR-fEPSPs traces are the average of 60 successive sweeps before (1) and during (2) bath ap- plications of D-serine and then (3) unclustered ephrinB3-Fc. The effect of unclustered ephrinB3-Fc was occluded by previous application of exogenous D-serine (50 µM). (E) Each connected circle represents paired mean NMDAR-fEPSP slope, for each individual experiment averaged in (D) measured in D-serine condition before (20–40 min) and after unclustered ephrinB3-Fc application (110–130 min), relative to baseline (-20–0 min). n.s. (P > 0.05). Wilcoxon signed-rank test.

Article Snippet: Langlais et al. Progress in Neurobiology 248 (2025) 102747 under agitation with an AffiniPure goat anti-human IgG1-Fc fragment specific antibody (1:2; Jackson ImmunoResearch).

Techniques: Activity Assay, Inhibition, MANN-WHITNEY, Two Tailed Test

Journal: Progress in neurobiology

Article Title: Astrocytic EphB3 receptors regulate d-serine-gated synaptic plasticity and memory.

doi: 10.1016/j.pneurobio.2025.102747

Figure Lengend Snippet: Fig. 5. EphB3-ephrinB3 interaction regulates LTP induction. (A), (D), Representative fEPSPs traces are the average of 30 successive sweeps before (1) and after (2) high frequency stimulation (HFS; 3 ×100 Hz, 1 sec, 20 sec interval; arrows). Slices were incubating 1 h with either unclustered IgG1-Fc (1.08 µg/ml) or ephrinB3-Fc (2.0 µg/ml), and 20 min with (D) or without (A) D-serine (50 µM), before HFS.(B), (E) Bar graphs representing the post-tetanic potentiation (PTP) measured the first minute after the induction. Results are presented as means ± s.e.m. for fEPSP slopes measured just after HFS (0–1 min) relative to baseline (-10–0 min). While the PTP was not altered (B), the LTP (A, C) was impaired when the endogenous interaction of EphB3 receptor with its endogenous ligand was decreased using unclustered ephrinB3-Fc.(C), (F) Bar graph summarizing the potentiation of synaptic transmission in (A) and (D). Results are presented as means ± s.e.m. for fEPSP slopes measured after HFS (50–60 min) relative to baseline (-10–0 min). ** P < 0.01. Mann Whitney test, two-tailed.

Article Snippet: Langlais et al. Progress in Neurobiology 248 (2025) 102747 under agitation with an AffiniPure goat anti-human IgG1-Fc fragment specific antibody (1:2; Jackson ImmunoResearch).

Techniques: Transmission Assay, MANN-WHITNEY, Two Tailed Test

Journal: mAbs

Article Title: RUBY® - a tetravalent (2+2) bispecific antibody format with excellent functionality and IgG-like stability, pharmacology and developability properties.

doi: 10.1080/19420862.2024.2330113

Figure Lengend Snippet: Figure 1. RUBY format structure. RUBY bsAbs comprise of an IgG (dark green, light green and gray) coupled to two fabs (dark and light turquoise). Three separate chains make up a construct, (1) a long chain that consists of the IgG heavy chain (dark green and gray) and the light chain of the additional fab fragment (light turquoise), (2) a light chain (light green) that binds to the VH and CH1 domains of the IgG part, and (3) a short heavy chain (dark turquoise) that binds to the light chain appended to the IgG.

Article Snippet: HRP conjugated goat anti-human IgG1 Fc (0.004 μg/ml, #109-035-098, Jackson ImmunoResearch) was used as detection in the mono ELISA and biotinylated human Fc conjugated human EpCAM (0.2 μg/ml, #10694- H02H-B, Sino Biological) followed by HRP conjugated streptavidin (#21126, Pierce) were used as detection in the dual ELISA.

Techniques: Construct

Journal: mAbs

Article Title: RUBY® - a tetravalent (2+2) bispecific antibody format with excellent functionality and IgG-like stability, pharmacology and developability properties.

doi: 10.1080/19420862.2024.2330113

Figure Lengend Snippet: Figure 4. RUBY bsAbs interaction with human FcRn and pharmacokinetics. BLI binding kinetics measurements of (a) RUBY bsAb and (b) monoclonal antibody control displaying similar interaction with human FcRn. (c) C57bl/6 mice were intravenously (i.v.) administered with molar equivalent doses of a human IgG1 LALA-mutated RUBY v9 bsAb (167 µg) or a corresponding mAb control (100 µg). Blood was collected 1, 2, 4, 24, 72 and 168 hours post dosing (n = 3/timepoint) and serum concentration of human IgG was measured using a human heavy chain-specific ELISA. (d) Human CD40 transgenic mice were administered with molar equivalent doses i.v. of two CD40-TAA RUBY bsAbs (360 µg) and a CD40 mAb (216 µg). Blood was collected 0.5, 1, 2, 4, 6, 24, 48 and 72 hours post dosing (n = 3/timepoint) and serum concentration of human IgG was measured using a human heavy chain-specific ELISA. (e) CEA-CD40 RUBY bsAb #2 was administered i.v. at 10 or 37.5 mg/kg to one female and one male cynomolgus monkey per dose level. Blood was collected 0.08, 0.5, 2, 4, 8, 24, 48, 72, 96 and 168 hours post dosing and serum concentration of human IgG was measured using a human CD40 and human CEACAM5 dual target ELISA. The ratio of full RUBY molecules over total human IgG in the serum of (f) human CD40 transgenic mice following i.V. administration of 417 µg CD40-EpCAM RUBY v9 #2, or (g) cynomolgus monkeys administered 10–37.5 mg/kg CEA-CD40 RUBY bsAb #2 was determined using ELISA. The graphs show the mean (±SD) of 3 mice/timepoint (c, c, and f), measured values over time for individual animals (e) or the mean (±SD) of four animals (g).

Article Snippet: HRP conjugated goat anti-human IgG1 Fc (0.004 μg/ml, #109-035-098, Jackson ImmunoResearch) was used as detection in the mono ELISA and biotinylated human Fc conjugated human EpCAM (0.2 μg/ml, #10694- H02H-B, Sino Biological) followed by HRP conjugated streptavidin (#21126, Pierce) were used as detection in the dual ELISA.

Techniques: Drug discovery, Binding Assay, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Transgenic Assay

Journal: mAbs

Article Title: RUBY® - a tetravalent (2+2) bispecific antibody format with excellent functionality and IgG-like stability, pharmacology and developability properties.

doi: 10.1080/19420862.2024.2330113

Figure Lengend Snippet: Figure 5. Tumor localization and hydrodynamic sizes of RUBY bsAbs and controls. (a) RUBY v9 bsAb efficiently localized to the tumor area, where they bound EpCAM expressing tumor. Mice were inoculated s.c. with MB49 tumors expressing human EpCAM or the same cell line lacking the EpCAM expression. The mice received a single CD40-EpCAM RUBY v9 #2 bsAb, CD40 mAb or vehicle i.p. injection and 24 hours later the tumors were collected and the frequency of IgG+ cells were analyzed by flow cytometry. (b) A RUBY bsAb and (c) Monoclonal antibody control displayed similar hydrodynamic sizes as determined by DLS.

Article Snippet: HRP conjugated goat anti-human IgG1 Fc (0.004 μg/ml, #109-035-098, Jackson ImmunoResearch) was used as detection in the mono ELISA and biotinylated human Fc conjugated human EpCAM (0.2 μg/ml, #10694- H02H-B, Sino Biological) followed by HRP conjugated streptavidin (#21126, Pierce) were used as detection in the dual ELISA.

Techniques: Expressing, Injection, Flow Cytometry, Control