her2 monoclonal antibody  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: From byproduct to biotherapeutic: Comparative study of buffy coats and leukoreduction system chambers for NK cell-based immunotherapies

doi: 10.1016/j.isci.2026.114907

Figure Lengend Snippet: Functional characterization of NK cells (A) Degranulation capacity of NK cells following a 2 h co-culture with K562 leukemia cells, measured by surface CD107a expression ( n = 5). (B and C) Europium-based cytotoxicity assay assessing the specific lysis of K562 target cells at E:T ratios of 10:1, 5:1, 3:1, 1:1, and 0.5:1 on day 7 (B) and day 14 (C) of culture to monitor functional decline over time ( n = 6 for both time points). Here, all comparisons were not significant and are summarized as such (n.s.: p > 0.05). (D) IFN-γ release following 16 h of co-culture of NK cells with K562 cells, quantified via Luminex assay ( n = 5). (E) Kinetic live-cell killing assay using the Incucyte monitoring of Nuclight Red signal loss in K562 cells over 72 h at an E:T ratio of 1:1 ( n = 6). (F) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis thresholds during Incucyte co-culture with K562 cells. (G) CD107a degranulation assay of NK cells with or without pre-incubation with trastuzumab (anti-HER2 antibody) during a 2 h co-culture with MDA-MB-453 cells ( n = 5). (H) Incucyte-based cytotoxicity assay shows the real-time lysis of MDA-MB-453 cells by trastuzumab-loaded NK cells at an E:T ratio of 1:1 over 72 h ( n = 6). (I) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis during the ADCC assay. (n varies as not all samples reached the benchmark values within the observation period). All ADCC experiments have been performed after 14 days of in vitro NK cell expansion. N/A indicates that statistical analysis was not possible for this group. 9J) CD107a degranulation of unmodified NK cells and ErbB2-CAR-NK cells after 2 h co-culture with MDA-MB-453 cells ( n = 5). (K) Incucyte-based cytotoxicity assay showing killing dynamics of ErbB2-CAR-NK cells against MDA-MB-453 cells at an E:T ratio of 1:1 over 72 h ( n = 6). (L) Proportion of donors reaching 25%, 50%, and 75% cumulative lysis during the CAR-mediated cytotoxicity assay. (n varies as not all samples reached the benchmark values within the observation period). All CAR-killing experiments have been performed after 14 days of in vitro NK cell expansion. Unless otherwise stated, n refers to biologically independent donor samples and is indicated per group. Data are shown as mean ± SEM. p-values were determined by Student’s t test with Welsh correction (A, D, F, G, I, J, and L) or by one-way ANOVA (B and C) with Tukey post-test (K and L).

Article Snippet: Mouse Monoclonal anti-CD340 (Her2) , Miltenyi Biotec , Cat# 130-124-474.

Techniques: Functional Assay, Co-Culture Assay, Expressing, Cytotoxicity Assay, Lysis, Luminex, Degranulation Assay, Incubation, ADCC Assay, In Vitro

Journal: NPJ Precision Oncology

Article Title: Quantitative HER2 tissue and plasma profiling predicts the activity of trastuzumab deruxtecan for breast cancer

doi: 10.1038/s41698-026-01365-6

Figure Lengend Snippet: A TTNT with T-DXd by HER2 status of the disease; B TTNT with T-DXd by clinical subtype of the disease; C TTNT with T-DXd depending on changes in HER2 status (low vs. 0) between the primary tumor and last metastatic biopsy; D TTNT with T-DXd depending on changes in HER2 status (low vs. 0) between the first and most recent metastatic biopsy; E OS with T-DXd by HER2 status of the disease; F OS with T-DXd by clinical subtype of the disease. T-DXd, trastuzumab deruxtecan; TTNT, time to next treatment; HER2, human epidermal growth factor receptor 2; OS, overall survival; HR, hormone receptor; TNBC, triple-negative breast cancer.

Article Snippet: The Leica BOND Rx autostainer protocol is as follows: deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97 ° C for 20 min, blocking with ReadyProbes Endogenous HRP & AP blocking solution ( R37629 , Invitrogen) for 10 min and with BSA for 30 min, 1 h incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, #2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3, REF#M3515, Dako), amplification with Rabbit Envision+ System–HRP labeled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody (REF#A11003) for 1 h, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 min and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 min.

Techniques:

Journal: NPJ Precision Oncology

Article Title: Quantitative HER2 tissue and plasma profiling predicts the activity of trastuzumab deruxtecan for breast cancer

doi: 10.1038/s41698-026-01365-6

Figure Lengend Snippet: Samples are ordered by HER2 IHC subtype and TTNT from the start of T-DXd treatment. Clinical parameters displayed include HER2 IHC subtype (HER2-zero, HER2-low, HER2-positive), latest HER2 IHC prior to T-DXd, number of prior lines of chemotherapy, and receipt of sacituzumab govitecan before T-DXd. An asterisk on the TTNT bar indicates treatment is still ongoing. Bar plots show scores for each assay, and these values are also represented with color-coded squares, along with their corresponding quartiles for the HS-HER2 and RPPA HER2 assays. For HER2DX ERBB2, score groups are displayed based on the recommended cutoffs: Low (1–32), Med (33–50), and High (51–99) . Some cases are depicted as HER2-positive given prior HER2-positivity, despite the closest biopsy to T-DXd administration being formally HER2-negative (as suggested by the IHC score). HS-HER2, High Sensitivity-human epidermal growth factor receptor 2; RPPA, Reverse Phase Protein Array; T-DXd, trastuzumab deruxtecan; TTNT, time to next treatment. *did not experience TTNT event.

Article Snippet: The Leica BOND Rx autostainer protocol is as follows: deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97 ° C for 20 min, blocking with ReadyProbes Endogenous HRP & AP blocking solution ( R37629 , Invitrogen) for 10 min and with BSA for 30 min, 1 h incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, #2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3, REF#M3515, Dako), amplification with Rabbit Envision+ System–HRP labeled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody (REF#A11003) for 1 h, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 min and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 min.

Techniques: Protein Array

Journal: NPJ Precision Oncology

Article Title: Quantitative HER2 tissue and plasma profiling predicts the activity of trastuzumab deruxtecan for breast cancer

doi: 10.1038/s41698-026-01365-6

Figure Lengend Snippet: A Workflow for the quantification of HS-HER2; B TTNT with T-DXd according to HS-HER2 quartiles; C OS with T-DXd according to HS-HER2 quartiles; D TTNT with T-DXd according to HS-HER2 median (HER2-positive disease only); E TTNT with T-DXd in the HS-HER2 cohort according to the traditional HER2 IHC classification of HER2-positive, HER2-low-HER2-0 in the HS-HER2 population. T-DXd trastuzumab deruxtecan, TTNT time to next treatment, HS-HER2 High Sensitivity-HER2, IHC immunohistochemistry, ROI region of interest, OS overall survival.

Article Snippet: The Leica BOND Rx autostainer protocol is as follows: deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97 ° C for 20 min, blocking with ReadyProbes Endogenous HRP & AP blocking solution ( R37629 , Invitrogen) for 10 min and with BSA for 30 min, 1 h incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, #2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3, REF#M3515, Dako), amplification with Rabbit Envision+ System–HRP labeled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody (REF#A11003) for 1 h, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 min and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 min.

Techniques: Immunohistochemistry

Journal: NPJ Precision Oncology

Article Title: Quantitative HER2 tissue and plasma profiling predicts the activity of trastuzumab deruxtecan for breast cancer

doi: 10.1038/s41698-026-01365-6

Figure Lengend Snippet: A Workflow for the CLIA-based RPPA analysis; B TTNT with T-DXd according to RPPA quantified total HER2 protein quartiles; C . OS with T-DXd according to RPPA HER2 protein expression quartiles; D TTNT with T-DXd according to RPPA measured HER2 activation (phosphoHER2 Y1248) quartiles; E OS with T-DXd according to RPPA measured HER2 activation (phosphoHER2 Y1248) quartiles; T-DXd trastuzumab deruxtecan, CLIA-RPPA Clinical Laboratory Improvement Amendments- Reverse Phase Protein Array, TTNT time to next treatment, OS overall survival.

Article Snippet: The Leica BOND Rx autostainer protocol is as follows: deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97 ° C for 20 min, blocking with ReadyProbes Endogenous HRP & AP blocking solution ( R37629 , Invitrogen) for 10 min and with BSA for 30 min, 1 h incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, #2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3, REF#M3515, Dako), amplification with Rabbit Envision+ System–HRP labeled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody (REF#A11003) for 1 h, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 min and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 min.

Techniques: Expressing, Activation Assay, Protein Array

Journal: NPJ Precision Oncology

Article Title: Quantitative HER2 tissue and plasma profiling predicts the activity of trastuzumab deruxtecan for breast cancer

doi: 10.1038/s41698-026-01365-6

Figure Lengend Snippet: A TTNT with T-DXd according to RPPA measured TOPO1 protein expression median (HER2-negative only); B OS with T-DXd according to RPPA measured TOPO1 protein expression median (HER2-negative only); C TTNT with T-DXd according to RPPA Trop2 quartiles; D TTNT with T-DXd according to RPPA measured EGFR protein expression quartiles; E TTNT with T-DXd according to RPPA measured phosphoHER3 protein expression quartiles; F TTNT with T-DXd according to the traditional HER2 IHC classification of HER2-positive, HER2-low-HER2-0 in the RPPA population. T-DXd trastuzumab deruxtecan, TTNT time to next treatment, RPPA Reverse Phase Protein Array, OS overall survival.

Article Snippet: The Leica BOND Rx autostainer protocol is as follows: deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97 ° C for 20 min, blocking with ReadyProbes Endogenous HRP & AP blocking solution ( R37629 , Invitrogen) for 10 min and with BSA for 30 min, 1 h incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, #2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3, REF#M3515, Dako), amplification with Rabbit Envision+ System–HRP labeled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody (REF#A11003) for 1 h, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 min and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 min.

Techniques: Expressing, Protein Array

Journal: NPJ Precision Oncology

Article Title: Quantitative HER2 tissue and plasma profiling predicts the activity of trastuzumab deruxtecan for breast cancer

doi: 10.1038/s41698-026-01365-6

Figure Lengend Snippet: A Description of HER2DX gene expression modules; association between the HER2 amplicon module with TTNT ( B ) and OS in all patients ( C ); association between the HER2 amplicon module with TTNT ( D ) and OS in HER2-positive MBC ( E ); F association between TTNT and the luminal module in patients with HER2-negative disease; G TTNT with T-DXd according to the traditional HER2 IHC classification of HER2-positive, HER2-low-HER2-0 in the HER2DX population. TTNT time to next treatment, OS overall survival, MBC metastatic breast cancer, T-DXd trastuzumab deruxtecan, IHC immunohistochemistry.

Article Snippet: The Leica BOND Rx autostainer protocol is as follows: deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97 ° C for 20 min, blocking with ReadyProbes Endogenous HRP & AP blocking solution ( R37629 , Invitrogen) for 10 min and with BSA for 30 min, 1 h incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, #2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3, REF#M3515, Dako), amplification with Rabbit Envision+ System–HRP labeled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody (REF#A11003) for 1 h, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 min and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 min.

Techniques: Gene Expression, Amplification, Immunohistochemistry

Journal: NPJ Precision Oncology

Article Title: Quantitative HER2 tissue and plasma profiling predicts the activity of trastuzumab deruxtecan for breast cancer

doi: 10.1038/s41698-026-01365-6

Figure Lengend Snippet: A Description of DNADX workflow; B Outcomes with T-DXd for metastatic breast cancer according to DNADX-detected tumor fraction; C Outcomes with T-DXd for metastatic breast cancer according to DNADX subtype; D Outcomes with T-DXd for metastatic breast cancer according to the DNADX HER2 signature; E Outcomes with T-DXd for metastatic breast cancer according to HER2 IHC status in the cohort of patients with detectable tumor fraction. T-DXd trastuzumab deruxtecan, TTNT, time to next treatment, OS overall survival.

Article Snippet: The Leica BOND Rx autostainer protocol is as follows: deparaffinization with BOND dewax solution (AR9222), antigen retrieval with BOND HIER epitope retrieval solution 2 (AR9640) at 97 ° C for 20 min, blocking with ReadyProbes Endogenous HRP & AP blocking solution ( R37629 , Invitrogen) for 10 min and with BSA for 30 min, 1 h incubation with primary rabbit monoclonal HER2 antibody (clone 29D8, #2165, IgG, Cell Signaling) at optimal concentration of 1ug/ml mixed together with 1:100 concentration of pan-CK (Clones AE1/AE3, REF#M3515, Dako), amplification with Rabbit Envision+ System–HRP labeled polymer anti-Rabbit (K400311-2, Dako) mixed together with 1:100 dilution of a green-fluorescent Alexa Fluor 546 Goat-anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody (REF#A11003) for 1 h, staining with 1:50 dilution of a red-fluorescent Tyramide Signal Amplification (TSA) Cyanin 5 (SAT705A001EA, Akoya Biosciences) for 10 min and nuclear staining with 1:500 dilution of a blue-fluorescent 4′,6-diamidino-2-phenylindole (DAPI) for 10 min.

Techniques:

Journal: Frontiers in Oncology

Article Title: In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab

doi: 10.3389/fonc.2025.1747065

Figure Lengend Snippet: Fluorescence microscopy study of Trastuzumab–ICG binding to BT-20-Luc and HCC-1419-Luc cell lines. After 1 hour (1 h) and 6 hours (6 h) incubation, fluorescence microscopy results showed (A) no signal from the HER2-negative cell line (BT-20-Luc) and (B) weak signals at 1 h or strong signals at 6 h in the HER2-positive cell line (HCC-1419-Luc).

Article Snippet: Trastuzumab, a humanized monoclonal antibody targeting HER2, was obtained from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan).

Techniques: Fluorescence, Microscopy, Binding Assay, Incubation

Journal: Frontiers in Oncology

Article Title: In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab

doi: 10.3389/fonc.2025.1747065

Figure Lengend Snippet: Fluorescence microscopy study of Trastuzumab–FITC binding to BT-20-Luc and HCC-1419-Luc cell lines. After 0.5 h and 1 h incubation, fluorescence microscopy results showed (A) weak signals from the HER2-negative cell line (BT-20-Luc) and (B) strong signals in the HER2-positive cell line (HCC-1419-Luc).

Article Snippet: Trastuzumab, a humanized monoclonal antibody targeting HER2, was obtained from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan).

Techniques: Fluorescence, Microscopy, Binding Assay, Incubation

Journal: Frontiers in Oncology

Article Title: In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab

doi: 10.3389/fonc.2025.1747065

Figure Lengend Snippet: Flow cytometry analysis of BT-20-Luc and HCC-1419-Luc cell lines incubated with Trastuzumab–ICG. In vitro experiments showed (A) non-specific, weak binding to the HER2-negative cell line at 6 hours and (B) specific binding to the HER2-positive cell line at the same point.

Article Snippet: Trastuzumab, a humanized monoclonal antibody targeting HER2, was obtained from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan).

Techniques: Flow Cytometry, Incubation, In Vitro, Binding Assay

Journal: Frontiers in Oncology

Article Title: In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab

doi: 10.3389/fonc.2025.1747065

Figure Lengend Snippet: Flow cytometry analysis of BT-20-Luc and HCC-1419-Luc cell lines incubated with Trastuzumab–FITC. In vitro experiments showed (A) non-specific, weak binding to the HER2-negative cell line and (B) specific binding to the HER2-positive cell line.

Article Snippet: Trastuzumab, a humanized monoclonal antibody targeting HER2, was obtained from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan).

Techniques: Flow Cytometry, Incubation, In Vitro, Binding Assay

Journal: Frontiers in Oncology

Article Title: In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab

doi: 10.3389/fonc.2025.1747065

Figure Lengend Snippet: In vivo HER2-targeted imaging. (A) White image. (B) FLI was used to evaluate the binding of Trastuzumab–ICG. HER2-positive primary tumors and lymph node metastases exhibited higher fluorescence than HER2-negative lesions, consistent with activation of the quenched probe in HER2-positive tissues. (C) BLI confirmed tumor formation .In vivo imaging was performed in n = 5 mice, each bearing one HER2-positive and one HER2-negative tumor with their corresponding lymph nodes.

Article Snippet: Trastuzumab, a humanized monoclonal antibody targeting HER2, was obtained from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan).

Techniques: In Vivo, Imaging, Binding Assay, Fluorescence, Activation Assay, In Vivo Imaging

Journal: Frontiers in Oncology

Article Title: In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab

doi: 10.3389/fonc.2025.1747065

Figure Lengend Snippet: Ex vivo imaging and quantitative analysis of excised tumors (Primary orthotopic breast tumors and lymph node metastatic lesions). (A) White light image (left) and fluorescence/white light fusion image (right) of representative tumors. The HER2-positive tumor exhibits a distinct fluorescent signal, whereas the HER2-negative tumor shows no obvious signal. (B) Quantitative fluorescence intensity of the tumors shown in (A) , demonstrating significantly higher signal in the HER2-positive group (p < 0.001). Plotted values represent raw fluorescence ROI intensities. In total, n = 10 HER2-positive tumors and n = 10 HER2-negative tumors were analyzed.

Article Snippet: Trastuzumab, a humanized monoclonal antibody targeting HER2, was obtained from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan).

Techniques: Ex Vivo, Imaging, Fluorescence

Journal: Frontiers in Oncology

Article Title: In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab

doi: 10.3389/fonc.2025.1747065

Figure Lengend Snippet: Comparison between tumors derived from HER2-positive and HER2-negative cells. (A) Comparison of tumor size between groups revealed no significant difference. (B) Quantification of bioluminescence signal normalized by tumor volume; no statistically significant difference was detected. Plotted BLI values are normalized (radiance per mm 3 ). Analyses were performed on n = 10 HER2-positive tumors and n = 10 HER2-negative tumors.

Article Snippet: Trastuzumab, a humanized monoclonal antibody targeting HER2, was obtained from Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan).

Techniques: Comparison, Derivative Assay