Anti-IGF1 ADCC Enhanced Antibody (BI 836845) is an ADCC enhanced antibody produced by our Afuco™ platform. BI 836845 is a HumAb of the IgG1 isotype against the human insulin-like growth factors IGF-1 and IGF-2, which are up-regulated in many tumors. BI 836845 is well tolerated in patients with advanced solid tumours. An RBD of 1000 mg weekly was identified. An expansion phase is ongoing and recruiting patients with Ewing sarcoma or all tumor types with biopsiable disease.
Figure 1 Concentration of mouse IGF-1 in column fractions as measured by ELISA.
A, Mouse IGF-1 concentration in the flow-through fraction detected by ELISA with no pretreatment (black) or following an 80.6°C pretreatment (white) according to initial BI 836845 or control antibody concentration. B, The percentage of IGF-1 measured by ELISA (with 80.6°C pretreatment) in flow-through (white), first-elution (striped), and second-elution (dotted) fractions with respect to total fraction (black) after column processing. Error bars indicate SD of measurements from at least 2 independent column-processing experiments. CTL, control.
Mireuta, M., Birman, E., Barmash, M., & Pollak, M. (2014). Quantification of binding of IGF-1 to BI 836845, a candidate therapeutic antibody against IGF-1 and IGF-2, and effects of this antibody on IGF-1: IGFBP-3 complexes in vitro and in male C57BL/6 mice. Endocrinology, 155(3), 703-715.
Figure 2 Mouse serum levels of free IGFBP-3
A, Samples of undiluted serum containing different concentrations of BI 836845 or 300 μg/mL control IgG1 were incubated in microplate wells coated with IGFBP-3-specific antibody. Molecular species bound to the anti-IGFBP-3-coated wells were eluted in an acidic condition (100 mM glycine, pH 2.3), neutralized, and amounts of IGF-1 (black) or IGFBP-3 (white) were quantified by ELISA. Error bars represent SD of measurements from at least 2 independent experiments. B, Schematic representation of results from Figure 2 illustrating the reciprocity of IGF-1-IGFBP (white squares) or IGF-1:BI 836845 (black squares) complexes. As the concentration of BI 836845 increases, the proportion of IGF-1 bound to IGFBP-3 decreases. The concentration of IGFBP-3 in mouse serum is approximately 20 nM. CTL, control.
Mireuta, M., Birman, E., Barmash, M., & Pollak, M. (2014). Quantification of binding of IGF-1 to BI 836845, a candidate therapeutic antibody against IGF-1 and IGF-2, and effects of this antibody on IGF-1: IGFBP-3 complexes in vitro and in male C57BL/6 mice. Endocrinology, 155(3), 703-715.
Figure 3 Mouse serum levels of BI 836845, total IGF-1, and potentially bioactive IGF-1 (ie, non-BI 836845-complexed IGF-1)
A, BI 836845 concentration assayed by ELISA in serum from C57 BL/6 mice treated with indicated doses of BI 836845 for 2 weeks, administered twice weekly, and humanely destroyed 4 days after the last injection. B, IGF-1 concentration in the same sera as in panel A. C, IGF-1 concentration in the flow-through fraction after Protein-A agarose column separation of sera used for panel A. D, GH concentration in the same sera as in panel A.
Mireuta, M., Birman, E., Barmash, M., & Pollak, M. (2014). Quantification of binding of IGF-1 to BI 836845, a candidate therapeutic antibody against IGF-1 and IGF-2, and effects of this antibody on IGF-1: IGFBP-3 complexes in vitro and in male C57BL/6 mice. Endocrinology, 155(3), 703-715.
Figure 4 Mouse serum levels of BI 836845, total IGF-1, and potentially bioactive IGF-1 (ie, non-BI 836845-complexed IGF-1)
A, BI 836845 concentration assayed by ELISA in total serum from C57 BL/6 mice treated once with 100 mg/kg BI 836845 and humanely destroyed at various time points over 48 hours. B, IGF-1 concentration in the same sera as in panel A. C, IGF-1 concentration in the flow-through fraction after Protein-A agarose column separation of sera used for panel A. D, Data from panels A, B, and C represented in molar form. Binding sites are obtained by a doubling of BI 836845 concentrations because 2 binding sites per antibody are present.
Mireuta, M., Birman, E., Barmash, M., & Pollak, M. (2014). Quantification of binding of IGF-1 to BI 836845, a candidate therapeutic antibody against IGF-1 and IGF-2, and effects of this antibody on IGF-1: IGFBP-3 complexes in vitro and in male C57BL/6 mice. Endocrinology, 155(3), 703-715.
Figure 5 Mouse plasma levels of IGFBP-3 and ALS
A, IGFBP-3 concentration assayed by ELISA in plasma from C57 BL/6 mice treated once with 100 mg/kg BI 836845 (black) or vehicle (white) over 48 hours. In treated set, error bars represent SD of measurements from n = 13 mice for the 0 time point, n = 12 for the 2-hour time point, n = 9 for the 6-hour time point, n = 6 for the 24-hour time point, and n = 3 for the 48 hours time point. In vehicle set, n = 4 at all time points. Global P value is less than 0.01 for repeated-measures ANOVA; *, P < .05; **, P < .01 with respect to control at specific time points using a Bonferroni correction for multiple testing. B, ALS concentration in plasma samples from a single group of 3 mice measured by Western blot sequentially at 0, 2, 6, 24, and 48 hours after BI 836845 treatment and in a second group of 3 mice measured sequentially at 0, 2, and 6 hours. Below each blot are identical measurements and time points from a group of 3 mice having received vehicle treatment.
Mireuta, M., Birman, E., Barmash, M., & Pollak, M. (2014). Quantification of binding of IGF-1 to BI 836845, a candidate therapeutic antibody against IGF-1 and IGF-2, and effects of this antibody on IGF-1: IGFBP-3 complexes in vitro and in male C57BL/6 mice. Endocrinology, 155(3), 703-715.
Figure 6 Mouse plasma levels of GH and mouse liver mRNA levels of IGF-1, IGFBP-3, and ALS
A, GH concentration assayed by ELISA in plasma from C57 BL/6 mice treated once with 100 mg/kg BI 836845 (black) or vehicle (white) over 48 hours. In treated set, error bars represent SD of measurements from n = 13 mice for the 0 time point, n = 12 for the 2-hour time point, n = 9 for the 6-hour time point, n = 6 for the 24-hour time point, and n = 3 for the 48-hour time point. In vehicle set, n = 4 at all time points. Global P value is less than .05 for repeated-measures ANOVA; *, P < .05 with respect to control at specific time points using a Bonferroni correction for multiple testing. B, GH concentration assayed at various time points by ELISA in plasma from C57 BL/6 mice treated daily with 100 mg/kg BI 836845 (black) or vehicle (white). Error bars represent SD of measurements from n = 3 mice for all time points. Global P value is < .0001 for repeated-measures ANOVA; **, P < .01; ***, P < .001 and with respect to time 0 using a Bonferroni correction. C, IGF-1 mRNA levels were quantified in livers of mice humanely destroyed at different time points following a single injection of 100 mg/kg BI 836845 with respect to total cDNA (white) or with respect to β-actin as a reference gene (white). The term "vehicle" refers to mice having received vehicle treatment and humanely destroyed at the end of the experiment, ie, 48 hours later.
Mireuta, M., Birman, E., Barmash, M., & Pollak, M. (2014). Quantification of binding of IGF-1 to BI 836845, a candidate therapeutic antibody against IGF-1 and IGF-2, and effects of this antibody on IGF-1: IGFBP-3 complexes in vitro and in male C57BL/6 mice. Endocrinology, 155(3), 703-715.
Figure 7 IGF pathway signal transduction in colorectal cancer cell lines.
Colorectal cancer cell lines were treated in vitro for 24 hours with BI 885578 (500 nmol/L) or an equivalent volume of DMSO (control) in fully supplemented culture medium. Phospho-IGF1R (A) and phospho-Akt (B) levels were measured in cell lysates using Bio-Plex assays. For each assay, mean values SDs are presented, which were determined from three or more replicates. Mean baseline phospho-IGF1R and phospho-Akt levels <50 U were considered below the detection limit for the assays and for these cell lines, the effects of BI 885578 treatment on those markers were not statistically evaluated. The cell lines are sorted according to IGF2 expression levels. IGF2high ¼ TPM 72. IGF2low ¼ TPM < 72; , P < 0.05 for treated versus control group. C, GEO and SW48 cells were treated for 10 minutes with or without IGF1 (100 ng/mL) in fully supplemented culture medium. Levels of the indicated proteins in cell lysates were analyzed by Western blotting. D, GEO cells were treated for 24 hours with or without BI 885578 (500 nmol/L) or the IGF1/2 antibody xentuzumab (BI 836845, 500 nmol/L) in fully supplemented culture medium before Western blotting analysis of lysates.
Sanderson, M. P., Hofmann, M. H., Garin-Chesa, P., Schweifer, N., Wernitznig, A., Fischer, S., ... & Arnhof, H. (2017). The IGF1R/INSR inhibitor BI 885578 selectively inhibits growth of IGF2-overexpressing colorectal cancer tumors and potentiates the efficacy of anti-VEGF therapy. Molecular cancer therapeutics, 16(10), 2223-2233.
Figure 8 Efficacy of the combination of BI 885578 with anti–VEGF-A in colorectal cancer xenograft models.
For each xenograft model, tumor-bearing mice (7/group) were treated with vehicle control, BI 885578 (40 mg/kg), anti-VEGF-A (15 mg/kg) or the combination of these agents. Relative tumor volume to baseline for each mouse and TGI data for each group are indicated for the last day of treatment. Data for only 6 CL-14 tumor-bearing mice are indicated in the BI 885578-treated group due to the unexplained mortality of one mouse on day 19. The indicated P values refer to the comparison of TGI data from the combination versus the highest single agent (HSA). IGF2 (high) = TPM ≧ 72. IGF2 (low) = TPM ≦ 72.
Sanderson, M. P., Hofmann, M. H., Garin-Chesa, P., Schweifer, N., Wernitznig, A., Fischer, S., ... & Arnhof, H. (2017). The IGF1R/INSR inhibitor BI 885578 selectively inhibits growth of IGF2-overexpressing colorectal cancer tumors and potentiates the efficacy of anti-VEGF therapy. Molecular cancer therapeutics, 16(10), 2223-2233.
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• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production
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CAT | Product Name | Application | Type |
---|---|---|---|
TAB-385CL | Anti-Human IGF1 Recombinant Antibody (BI 836845) | WB, FuncS | Antibody |
CAT | Product Name | Application | Type |
---|---|---|---|
NEUT-1072CQ | Mouse Anti-IGF1 Recombinant Antibody (clone M23) | Neut, FC, ICC, IF | Mouse IgG1, κ |
NEUT-1073CQ | Mouse Anti-IGF1 Recombinant Antibody (clone SPM406) | WB, Neut, FC, ICC, IF, IHC-P, IP | Mouse IgG1, κ |
NEUT-1074CQ | Hamster Anti-Igf1 Recombinant Antibody (clone CBL534) | WB, ELISA, Neut | Hamster IgG |
CAT | Product Name | Application | Type |
---|---|---|---|
MOR-1754 | Rabbit Anti-IGF1 Recombinant Antibody (clone DS1754AB) | WB | Rabbit IgG |
CAT | Product Name | Application | Type |
---|---|---|---|
HPAB-0770-CN | Human Anti-IGF1 Recombinant Antibody (HPAB-0770-CN) | WB, FC | Human IgG1 |
HPAB-0771-CN | Human Anti-IGF1 Recombinant Antibody (HPAB-0771-CN) | WB, FC | Human IgG |
HPAB-0038LY | Human Anti-IGF1 Recombinant Antibody (HPAB-0038LY) | WB, Inhib | Human IgG1 |
HPAB-0160-YJ | Human Anti-IGF1 Recombinant Antibody (HPAB-0160-YJ) | ELISA, Neut, FuncS | Human IgG1 |
HPAB-0161-YJ | Human Anti-IGF1 Recombinant Antibody (HPAB-0161-YJ) | ELISA, Neut, FuncS | Human IgG1 |
CAT | Product Name | Application | Type |
---|---|---|---|
HPAB-0770-CN-F(E) | Human Anti-IGF1 Recombinant Antibody; Fab Fragment (HPAB-0770-CN-F(E)) | WB, FC | Human Fab |
HPAB-0160-YJ-F(E) | Human Anti-IGF1 Recombinant Antibody; Fab Fragment (HPAB-0160-YJ-F(E)) | ELISA, Neut, FuncS | Human Fab |
HPAB-0161-YJ-F(E) | Human Anti-IGF1 Recombinant Antibody; Fab Fragment (HPAB-0161-YJ-F(E)) | ELISA, Neut, FuncS | Human Fab |
HPAB-0711-FY-S(P) | Mouse Anti-IGF1 Recombinant Antibody (clone AB010); Fab Fragment | ELISA | Mouse Fab |
HPAB-0556-YJ-F(E) | Mouse Anti-IGF1 Recombinant Antibody (clone CBI65YJ); Fab Fragment | ELISA | Mouse Fab |
CAT | Product Name | Application | Type |
---|---|---|---|
HPAB-646-FY-S(P) | Mouse Anti-IGF1 Recombinant Antibody (clone MuEM164); scFv Fragment | Inhib, FC | Mouse scFv |
HPAB-647-FY-S(P) | Human Anti-IGF1 Recombinant Antibody (clone huEM164 V1.0); scFv Fragment | Inhib, FC | Human scFv |
HPAB-0160-YJ-S(P) | Human Anti-IGF1 Recombinant Antibody; scFv Fragment (HPAB-0160-YJ-S(P)) | ELISA, Neut, FuncS | Human scFv |
HPAB-0161-YJ-S(P) | Human Anti-IGF1 Recombinant Antibody; scFv Fragment (HPAB-0161-YJ-S(P)) | ELISA, Neut, FuncS | Human scFv |
HPAB-0556-YJ-S(P) | Mouse Anti-IGF1 Recombinant Antibody (clone CBI65YJ); scFv Fragment | ELISA | Mouse scFv |
CAT | Product Name | Application | Type |
---|---|---|---|
MHC-CN0897 | PE-H-2Db/Human IGF1 (SHLVEALYL) MHC Tetramer | FCM, IHC | |
MHC-CN0898 | APC-H-2Db/Human IGF1 (SHLVEALYL) MHC Tetramer | FCM, IHC | |
MHC-CN0899 | FITC-H-2Db/Human IGF1 (SHLVEALYL) MHC Tetramer | FCM, IHC | |
MHC-CN0900 | H-2Db/Human IGF1 (SHLVEALYL) MHC Tetramer | FCM, IHC |
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