Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Evaluation of ON203, A Novel Bioengineered mAb Targeting Oxidized Macrophage Migration Inhibitory Factor as an Anticancer Therapeutic

doi: 10.1158/1535-7163.MCT-22-0676

Figure Lengend Snippet: Physicochemical characterization of the mAb variants revealed identical binding properties and reduced hydrophobicity of the bioengineered anti-oxMIF antibodies compared with C0008 (imalumab). A, Antibody specificity of oxMIF versus redMIF in solution using 50 ng/mL of an oxMIF surrogate (TNB-MIF) by ELISA. B, Antibody apparent affinity to human oxMIF immobilized to an ELISA plate. C, Antibody affinity to human and mouse oxMIF in solution (using ProClin300 to induce oxMIF structure) by ELISA. D, Real-time binding kinetics of immobilized ON103 and ON203 versus C0008 for soluble human and mouse oxMIF by SPR. Data are represented as mean ± SD of the calculated association constants [ka, 1/(mol/L s)], dissociation constants (kd, 1/second), and equilibrium constants (K D , nmol/L) of flow cells 2–4 (after the subtraction of background from the reference cell (FC1). E, HIC of bioengineered antibodies compared with imalumab (C0008). EC 50 , effective concentration at 50% signal.

Article Snippet: The first-generation human IgG1 anti-oxMIF mAb (imalumab) was identified from a large and highly diverse panel of 145 unique MIF-specific antibodies that were selected from Dyax Fab310 phage display human Fab library ( ).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Evaluation of ON203, A Novel Bioengineered mAb Targeting Oxidized Macrophage Migration Inhibitory Factor as an Anticancer Therapeutic

doi: 10.1158/1535-7163.MCT-22-0676

Figure Lengend Snippet: ON203 is efficacious in a prophylactic prostate cancer xenograft model. A, Schematic presentation of the prophylactic prostate cancer xenograft model created with www.biorender.com . B, Tumor volume of subcutaneous PC3 tumor-bearing mice treated 15 times every other day intraperitoneally with vehicle, 20 mg/kg isotype IgG control antibody, 20 mg/kg C0008, 5 mg/kg ON203, or 20 mg/kg ON203 followed for 42 days after randomization ( n = 9/treatment group). Median tumor volumes with interquartile ranges are shown. Levels of significance were calculated for all groups by ANOVA with Dunnett multiple comparisons test on log-transformed data; ON203 20 mg/kg versus vehicle + isotype IgG 20 mg/kg (black stars), ON203 20 mg/kg versus C0008 20 mg/kg (gray star) *, P < 0.05; **, P < 0.01. C, Relative change in animal body weight (%) from day 0 ( n = 9/treatment group, mean ± SEM). D, Representative immunostaining images of 20 mg/kg isotype IgG-, C0008-, or ON203-treated PC3 tumors; scale bar, 100 μm (CD31 and H&E) or 40 μm (Ki67). Top: CD31 immunostaining (red arrows highlight intravascular tumor cells, and black arrows mark CD31-positive blood vessels); middle: H&E; bottom: Ki67 immunostaining. E, Vessel density in non-necrotic areas quantified on CD31-stained IHC slides from 20 mg/kg isotype IgG control-, C0008-, or ON203-treated tumors ( n = 3/treatment group, mean ± SEM). F, Percentage of Ki67-positive cells quantified on Ki67-positive stained IHC slides from 20 mg/kg isotype IgG control-, C0008-, or ON203-treated tumors ( n = 3/treatment group, mean ± SEM). Levels of significance were calculated by one-sided Student t test, *, P <0.05. i.p., intraperitoneal; SEM, standard error of mean.

Article Snippet: The first-generation human IgG1 anti-oxMIF mAb (imalumab) was identified from a large and highly diverse panel of 145 unique MIF-specific antibodies that were selected from Dyax Fab310 phage display human Fab library ( ).

Techniques: Control, Transformation Assay, Immunostaining, Staining

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Evaluation of ON203, A Novel Bioengineered mAb Targeting Oxidized Macrophage Migration Inhibitory Factor as an Anticancer Therapeutic

doi: 10.1158/1535-7163.MCT-22-0676

Figure Lengend Snippet: ON203 inhibits tumor growth in a therapeutic prostate cancer xenograft model. A, Schematic presentation of the therapeutic prostate cancer xenograft model created with www.biorender.com . B, Tumor volume of subcutaneous PC3 tumor-bearing mice treated 15 times every other day intraperitoneally with vehicle, 60 mg/kg isotype IgG control antibody, 60 mg/kg C0008, or 20, 40, or 60 mg/kg ON203 followed for 34 days after randomization ( n = 7–9/treatment group). Mean tumor volumes ±SEM are shown. Only data up to day 29 could be evaluated, because from this point on several animals had to be sacrificed because of excessive tumor growth. Levels of significance were calculated for all groups by repeated measures two-way ANOVA with Dunnett multiple comparison correction on log-transformed data; ON203 versus C0008 (gray stars); ON203 versus vehicle + isotype IgG (black stars); ON203 40 mg/kg versus ON203 20 mg/kg (green stars) *, P ≤0.05; **, P < 0.01; ***, P < 0.001. C, Relative body weight change of animals in percent to day 0 ( n = 7–9/treatment group, mean ± SEM). D, Mean tumor growth inhibition (TGI, mean ± SEM) by C0008 (60 mg/kg) versus ON203 (20, 40, or 60 mg/kg) calculated in percentage relative to the mean tumor volume from vehicle plus isotype IgG. Levels of significance were calculated by ANOVA with Dunnett multiple comparisons test, *, P < 0.05. E, Representative images of Ki67 immunostaining of 60 mg/kg isotype IgG, 60 mg/kg C0008, or 40 mg/kg or 60 mg/kg ON203-treated PC3 tumors; scale bar, 60 μm. F, Percentage of Ki67-positive cells on IHC slides from 60 mg/kg isotype IgG, 60 mg/kg C0008, or 40 and 60 mg/kg ON203-treated tumors ( n = 6/treatment group, mean ± SEM). Levels of significance were calculated by one-sided Student t test on log-transformed data, *, P ≤0.05. i.p., intraperitoneal; s.c., subcutaneous; SEM, standard error of mean; TGI, tumor growth inhibition.

Article Snippet: The first-generation human IgG1 anti-oxMIF mAb (imalumab) was identified from a large and highly diverse panel of 145 unique MIF-specific antibodies that were selected from Dyax Fab310 phage display human Fab library ( ).

Techniques: Control, Comparison, Transformation Assay, Inhibition, Immunostaining