Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Flow cytometry for HER2-positive and EGFR-positive cells in populations of SK-OV-3 and/or MDA-MB-468 cells SK-OV-3 or MDA-MB-468 cells or mixtures of these cells immunostained with (A) anti-HER2 Alexa Fluor 488-conjugated trastuzumab or (B) with anti-EGFR Alexa Fluor 647-conjugated panitumumab were analyzed by flow cytometry. The proportion of HER2-positive or EGFR-positive cells was quantified as the area under the relevant peak for SK-OV-3 or MDA-MB-468 cells, respectively and plotted vs. the percentage of (C) SK-OV-3 cells or (D) MDA-MB-468 cells in the cell population.

Article Snippet: Tumors were embedded into formalin-fixed, paraffin blocks and 10 μm sections were immunostained at the Pathology Research Program Laboratory at the University Health Network for EGFR using rabbit anti-human EGFR monoclonal antibodies (Cell Signaling Technology, Product #4267) and HER2 using rabbit anti-human HER2 monoclonal antibodies (ThermoFisher/Invitrogen, Product #SP3).

Techniques: Flow Cytometry

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Flow cytometry for EGFR-positive cells in dissociated tumors composed of MDA-MB-468 and/or SK-OV-3 cells Tumor xenografts in NOD/SCID mice were dissociated at 6 weeks post s.c. inoculation of tumor cells then the recovered cells were immunostained with anti-EGFR Alexa Fluor 647-conjugated panitumumab and analyzed by flow cytometry. (A) Based on gating thresholds, MDA-MB-468 tumors were assigned as 96.6% EGFR-positive cells and 3.4% EGFR-negative cells. (B) SK-OV-3 tumors were assigned as 7.8% EGFR-positive cells and 92.2% EGFR-negative cells. (C) Replicate tumors ( n = 3) formed by the inoculation of 70% MDA-MB-468 cells and 30% SK-OV-3 cells were assigned as 76.3%, 71.4%, and 68.6% EGFR-positive cells and 24.1%, 28.6%, and 31.4% EGFR-negative cells.

Article Snippet: Tumors were embedded into formalin-fixed, paraffin blocks and 10 μm sections were immunostained at the Pathology Research Program Laboratory at the University Health Network for EGFR using rabbit anti-human EGFR monoclonal antibodies (Cell Signaling Technology, Product #4267) and HER2 using rabbit anti-human HER2 monoclonal antibodies (ThermoFisher/Invitrogen, Product #SP3).

Techniques: Flow Cytometry

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Immunohistochemical (IHC) staining of tumors composed of SK-OV-3 and/or MDA-MB-468 cells for HER2-positive and EGFR-positive cells Representative sections of tumors formed by the inoculation of (A) SK-OV-3 cells or (B) MDA-MB-468 cells or (C) a mixture of 30% SK-OV-3 cells and 70% MDA-MB-468 cells at 6 weeks post-inoculation in NOD/SCID mice. Sections were stained with hematoxylin and eosin (H&E) or immunostained for HER2 or EGFR. Sections are shown at 4× or 20× amplification. (D) The percentage of cells immunopositive for HER2 or EGFR in replicate tumor sections was quantified by image analysis and is shown for SK-OV-3 or MDA-MB-468 tumors or tumors formed by inoculating a mixture of 30% SK-OV-3 cells and 70% MDA-MB-468 cells. Data shown are mean ± SD ( n = 3). Statistically significant differences (Student’s t test; p < 0.05) are indicated by the asterisks.

Article Snippet: Tumors were embedded into formalin-fixed, paraffin blocks and 10 μm sections were immunostained at the Pathology Research Program Laboratory at the University Health Network for EGFR using rabbit anti-human EGFR monoclonal antibodies (Cell Signaling Technology, Product #4267) and HER2 using rabbit anti-human HER2 monoclonal antibodies (ThermoFisher/Invitrogen, Product #SP3).

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, Amplification

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: PET/CT images of NOD-SCID mice with s.c. SK-OV-3 tumors with homogeneous HER2 expression or tumors with heterogeneous HER2 expression formed from a mixture of SK-OV-3 and MDA-MB-468 cells Representative images (supine position) of NOD-SCID mice with s.c. SK-OV-3 tumors on the right flank that homogeneously express HER2 (blue arrow) and tumors formed by the inoculation of a mixture of SK-OV-3 cells (30%) and MDA-MB-468 cells (70%) with heterogeneous HER2 and EGFR expression on the left flank (red arrow). Images were obtained at 24 h and 48 h p.i. of (A) [ 64 Cu]Cu-NOTA-trastuzumab Fab-PEG 24 -EGF, (B) [ 64 Cu]Cu-NOTA-trastuzumab Fab or (C) [ 64 Cu]Cu-NOTA-EGF. All images were adjusted to the same intensity and the intensity scale (ranging from 1.5% ID/g to 12.5% ID/g) is shown at the right of each set of images. Normal organ uptake is visible on the 24 h and 48 h p.i. images but is labeled on the 48 h p.i. images. L: liver; K: kidneys; Sp: spleen.

Article Snippet: Tumors were embedded into formalin-fixed, paraffin blocks and 10 μm sections were immunostained at the Pathology Research Program Laboratory at the University Health Network for EGFR using rabbit anti-human EGFR monoclonal antibodies (Cell Signaling Technology, Product #4267) and HER2 using rabbit anti-human HER2 monoclonal antibodies (ThermoFisher/Invitrogen, Product #SP3).

Techniques: Positron Emission Tomography-Computed Tomography, Expressing, Labeling

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: PET/CT images of NOD-SCID mice with s.c. MDA-MB-468 tumors with homogeneous EGFR expression or tumors with heterogeneous EGFR expression formed from a mixture of MDA-MB-468 and SK-OV-3 cells Representative images (supine position) of NOD-SCID mice with s.c. MDA-MB-468 tumors on the right flank that homogeneously express EGFR (blue arrow) and tumors formed by the inoculation of a mixture of MDA-MB-468 cells (70%) and SK-OV-3 cells (30%) with heterogeneous HER2 and EGFR expression on the left flank (red arrow). Images were obtained at 24 h and 48 h p.i. of (A) [ 64 Cu]Cu-NOTA-trastuzumab Fab-PEG 24 -EGF, (B) [ 64 Cu]Cu-NOTA-trastuzumab Fab or (C) [ 64 Cu]Cu-NOTA-EGF. All images were adjusted to the same intensity and the intensity scale (ranging from 1.5% ID/g to 12.5% ID/g) is shown at the right of each set of images. Normal organ uptake is visible on the 24 h and 48 h p.i. images but is labeled on the 48 h p.i. images. L: liver; K: kidneys; Sp: spleen.

Article Snippet: Tumors were embedded into formalin-fixed, paraffin blocks and 10 μm sections were immunostained at the Pathology Research Program Laboratory at the University Health Network for EGFR using rabbit anti-human EGFR monoclonal antibodies (Cell Signaling Technology, Product #4267) and HER2 using rabbit anti-human HER2 monoclonal antibodies (ThermoFisher/Invitrogen, Product #SP3).

Techniques: Positron Emission Tomography-Computed Tomography, Expressing, Labeling

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet:

Article Snippet: Tumors were embedded into formalin-fixed, paraffin blocks and 10 μm sections were immunostained at the Pathology Research Program Laboratory at the University Health Network for EGFR using rabbit anti-human EGFR monoclonal antibodies (Cell Signaling Technology, Product #4267) and HER2 using rabbit anti-human HER2 monoclonal antibodies (ThermoFisher/Invitrogen, Product #SP3).

Techniques: Recombinant, Expressing, Software, Positron Emission Tomography, Computed Tomography, Flow Cytometry

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Flow cytometry for HER2-positive and EGFR-positive cells in populations of SK-OV-3 and/or MDA-MB-468 cells SK-OV-3 or MDA-MB-468 cells or mixtures of these cells immunostained with (A) anti-HER2 Alexa Fluor 488-conjugated trastuzumab or (B) with anti-EGFR Alexa Fluor 647-conjugated panitumumab were analyzed by flow cytometry. The proportion of HER2-positive or EGFR-positive cells was quantified as the area under the relevant peak for SK-OV-3 or MDA-MB-468 cells, respectively and plotted vs. the percentage of (C) SK-OV-3 cells or (D) MDA-MB-468 cells in the cell population.

Article Snippet: Rabbit anti-human EGFR monoclonal antibody , Cell Signaling Technology , #4267.

Techniques: Flow Cytometry

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Flow cytometry for EGFR-positive cells in dissociated tumors composed of MDA-MB-468 and/or SK-OV-3 cells Tumor xenografts in NOD/SCID mice were dissociated at 6 weeks post s.c. inoculation of tumor cells then the recovered cells were immunostained with anti-EGFR Alexa Fluor 647-conjugated panitumumab and analyzed by flow cytometry. (A) Based on gating thresholds, MDA-MB-468 tumors were assigned as 96.6% EGFR-positive cells and 3.4% EGFR-negative cells. (B) SK-OV-3 tumors were assigned as 7.8% EGFR-positive cells and 92.2% EGFR-negative cells. (C) Replicate tumors ( n = 3) formed by the inoculation of 70% MDA-MB-468 cells and 30% SK-OV-3 cells were assigned as 76.3%, 71.4%, and 68.6% EGFR-positive cells and 24.1%, 28.6%, and 31.4% EGFR-negative cells.

Article Snippet: Rabbit anti-human EGFR monoclonal antibody , Cell Signaling Technology , #4267.

Techniques: Flow Cytometry

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: Immunohistochemical (IHC) staining of tumors composed of SK-OV-3 and/or MDA-MB-468 cells for HER2-positive and EGFR-positive cells Representative sections of tumors formed by the inoculation of (A) SK-OV-3 cells or (B) MDA-MB-468 cells or (C) a mixture of 30% SK-OV-3 cells and 70% MDA-MB-468 cells at 6 weeks post-inoculation in NOD/SCID mice. Sections were stained with hematoxylin and eosin (H&E) or immunostained for HER2 or EGFR. Sections are shown at 4× or 20× amplification. (D) The percentage of cells immunopositive for HER2 or EGFR in replicate tumor sections was quantified by image analysis and is shown for SK-OV-3 or MDA-MB-468 tumors or tumors formed by inoculating a mixture of 30% SK-OV-3 cells and 70% MDA-MB-468 cells. Data shown are mean ± SD ( n = 3). Statistically significant differences (Student’s t test; p < 0.05) are indicated by the asterisks.

Article Snippet: Rabbit anti-human EGFR monoclonal antibody , Cell Signaling Technology , #4267.

Techniques: Immunohistochemical staining, Immunohistochemistry, Staining, Amplification

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: PET/CT images of NOD-SCID mice with s.c. SK-OV-3 tumors with homogeneous HER2 expression or tumors with heterogeneous HER2 expression formed from a mixture of SK-OV-3 and MDA-MB-468 cells Representative images (supine position) of NOD-SCID mice with s.c. SK-OV-3 tumors on the right flank that homogeneously express HER2 (blue arrow) and tumors formed by the inoculation of a mixture of SK-OV-3 cells (30%) and MDA-MB-468 cells (70%) with heterogeneous HER2 and EGFR expression on the left flank (red arrow). Images were obtained at 24 h and 48 h p.i. of (A) [ 64 Cu]Cu-NOTA-trastuzumab Fab-PEG 24 -EGF, (B) [ 64 Cu]Cu-NOTA-trastuzumab Fab or (C) [ 64 Cu]Cu-NOTA-EGF. All images were adjusted to the same intensity and the intensity scale (ranging from 1.5% ID/g to 12.5% ID/g) is shown at the right of each set of images. Normal organ uptake is visible on the 24 h and 48 h p.i. images but is labeled on the 48 h p.i. images. L: liver; K: kidneys; Sp: spleen.

Article Snippet: Rabbit anti-human EGFR monoclonal antibody , Cell Signaling Technology , #4267.

Techniques: Positron Emission Tomography-Computed Tomography, Expressing, Labeling

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet: PET/CT images of NOD-SCID mice with s.c. MDA-MB-468 tumors with homogeneous EGFR expression or tumors with heterogeneous EGFR expression formed from a mixture of MDA-MB-468 and SK-OV-3 cells Representative images (supine position) of NOD-SCID mice with s.c. MDA-MB-468 tumors on the right flank that homogeneously express EGFR (blue arrow) and tumors formed by the inoculation of a mixture of MDA-MB-468 cells (70%) and SK-OV-3 cells (30%) with heterogeneous HER2 and EGFR expression on the left flank (red arrow). Images were obtained at 24 h and 48 h p.i. of (A) [ 64 Cu]Cu-NOTA-trastuzumab Fab-PEG 24 -EGF, (B) [ 64 Cu]Cu-NOTA-trastuzumab Fab or (C) [ 64 Cu]Cu-NOTA-EGF. All images were adjusted to the same intensity and the intensity scale (ranging from 1.5% ID/g to 12.5% ID/g) is shown at the right of each set of images. Normal organ uptake is visible on the 24 h and 48 h p.i. images but is labeled on the 48 h p.i. images. L: liver; K: kidneys; Sp: spleen.

Article Snippet: Rabbit anti-human EGFR monoclonal antibody , Cell Signaling Technology , #4267.

Techniques: Positron Emission Tomography-Computed Tomography, Expressing, Labeling

Journal: iScience

Article Title: Bispecific radioimmunoconjugates exploit receptor heterogeneity for positron emission tomography of tumors expressing HER2 and/or EGFR

doi: 10.1016/j.isci.2024.109750

Figure Lengend Snippet:

Article Snippet: Rabbit anti-human EGFR monoclonal antibody , Cell Signaling Technology , #4267.

Techniques: Recombinant, Expressing, Software, Positron Emission Tomography, Computed Tomography, Flow Cytometry