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Image Search Results
Journal: EBioMedicine
Article Title: A novel HER2-targeting antibody 5G9 identified by large-scale trastuzumab-based screening exhibits potent synergistic antitumor activity
doi: 10.1016/j.ebiom.2020.102996
Figure Lengend Snippet: Characterization of trastuzumab-optimal-synergistic-epitope (TOSE) -binding monoclonal antibodies. (a) Schematic workflow of trastuzumab-based synergistic functional screening of TOSE-binding antibodies (4H2, 4C9, 4G6, 5F12 and 5G9). 1st screening: ELISA binding and competitive ELISA with trastuzumab or pertuzumab; 2nd screening: binding to HER2-positive cells; 3rd screening: trastuzumab-based cell proliferation inhibition assay. (b) Five monoclonal antibodies (4H2, 4C9, 4G6, 5F12 and 5G9) showed trastuzumab-synergistic bioactivity higher than pertuzumab in the BT-474 cell proliferation inhibition assay. TRA: trastuzumab; PER: pertuzumab. (c) The antibody epitope grouping assay was performed by competitive ELISA. Five monoclonal antibodies (4H2, 4C9, 4G6, 5F12 and 5G9) bound to a unique epitope-TOSE, which was different from both the trastuzumab-binding epitope and pertuzumab-binding epitope. (d) The binding specificity and species cross-reactivity were determined by ELISA. 5G9 bound to HER2 protein, but not to other EGFR family members, including EGFR, HER3 and HER4. (e) The cross-species binding reactivity was determined by ELISA. 5G9 was able to bind cynomolgus HER2 protein but not murine HER2 protein. The BT-474 cell proliferation inhibition assay, antibody epitope grouping assay and the cross-reactivity assay were repeated three times and the results were shown as the mean ± SD ( n = 3). Two-way ANOVA, *** P< 0.001.
Article Snippet: The recombinant proteins huEGFR (Cat# 10001-H02H), huHER2 (Cat# 10004-H02H), huHER3 (Cat# 10201-H02H), huHER4 (Cat# 10363-H02H), rhesus HER2 (Cat# 90020-K02H) and
Techniques: Binding Assay, Functional Assay, Enzyme-linked Immunosorbent Assay, Competitive ELISA, Inhibition
Journal: EBioMedicine
Article Title: A novel HER2-targeting antibody 5G9 identified by large-scale trastuzumab-based screening exhibits potent synergistic antitumor activity
doi: 10.1016/j.ebiom.2020.102996
Figure Lengend Snippet: 5G9 and trastuzumab synergistically inhibit HER2-positive cancer cell growth. (a) Inhibition of BT-474 cell viability was examined by Cell Titer Glo after 6 days of antibody treatment. The initial antibody concentration was 100 nM for both the monoclonal antibody and the antibody combination (1:1 ratio). Growth inhibition was calculated as the fraction of viable cells in the antibody treated group compared with untreated control cells. The cell proliferation inhibition assay was repeated three times and the results were shown as the mean ± SD ( n = 3). Two-way ANOVA, ** P< 0.01. TRA: trastuzumab; PER: pertuzumab. (b) Dose-effect plots and combination index (CI.) plots of the two combinations were calculated by using the method of Chou and Talalay with the commercial software Calcusyn. C.I. values for effective doses at which 50%, 75%, and 90% (ED 50 , ED 75 , and ED 90 , respectively) of cells were killed. Drug synergy was defined by C.I. values less than 1, the lower C.I. value, the more synergy. (c) Representative images from the EdU assay of BT-474 cells. The viable cells were stained blue by DAPI (blue) and the proliferation cells were stained red by EdU (red).The EdU assay was repeated three times and the representative images were shown. (d) Human breast cancer cell lines (SK-BR-3, AU-565, MCF-7) and human gastric cell lines (NCI-N87) were treated with 100 nM antibodies for 3 or 6 days and cell viability was determined by Cell Titer Glo. The experiments were repeated three times and the cell viability is shown as the mean ± SD ( n = 3). The relative HER2 expression of cell lines was detected and analyzed by flow cytometry and FlowJo software.
Article Snippet: The recombinant proteins huEGFR (Cat# 10001-H02H), huHER2 (Cat# 10004-H02H), huHER3 (Cat# 10201-H02H), huHER4 (Cat# 10363-H02H), rhesus HER2 (Cat# 90020-K02H) and
Techniques: Inhibition, Concentration Assay, Software, EdU Assay, Staining, Expressing, Flow Cytometry
Journal: EBioMedicine
Article Title: A novel HER2-targeting antibody 5G9 identified by large-scale trastuzumab-based screening exhibits potent synergistic antitumor activity
doi: 10.1016/j.ebiom.2020.102996
Figure Lengend Snippet: 5G9 and trastuzumab enhance cell apoptosis, HER2 internalization and disruption. (a) BT-474 cells were treated with 100 nM antibodies for 3 days. The number of viable cells was detected by Trypan Blue, and annexin-V and PI were used to stain early apoptotic cells and dead cells. TRA: trastuzumab; PER: pertuzumab. (b) BT-474 cells were treated with 100 nM antibodies for 3 days, and cell lysates were obtained and immunoblotted with a poly (ADP-ribose) polymerase (PARP) polyclonal antibody. Cleaved PARP is represented by 89 KD-fragments and full-length PARP (116 KD). (c) HER2 internalization induced by 5G9, trastuzumab or pertuzumab in SK-BR-3 cells. (d) The effect of trastuzumab on the internalization of 5G9 or pertuzumab in SK-BR-3 cells. FITClabeled 5G9 or pertuzumab was detected and analyzed by flow cytometry and FlowJo software. (e) The effect of 5G9 or pertuzumab on the internalization of trastuzumab in SK-BR-3 cells. APC labeled trastuzumab was detected and analyzed by flow cytometry and FlowJo software. (f) Effect of HER2 single agents or combination on receptor phosphorylation and downstream signaling cascade. BT-474 cells were treated with 100 nM antibodies for 3 days, and cell lysates were obtained and immunoblotted with antibodies against EGFR, p-EGFR, HER2, p-HER2, HER3, p-HER3, AKT, p-AKT, ERK, p-ERK, and GAPDH. All experiments were repeated three times and the representative images and data were shown. Two-way ANOVA, * P< 0.05, *** P< 0.001.
Article Snippet: The recombinant proteins huEGFR (Cat# 10001-H02H), huHER2 (Cat# 10004-H02H), huHER3 (Cat# 10201-H02H), huHER4 (Cat# 10363-H02H), rhesus HER2 (Cat# 90020-K02H) and
Techniques: Staining, Flow Cytometry, Software, Labeling
Journal: Biosensors
Article Title: Dual-Oriented Targeted Nanostructured SERS Label-Free Immunosensor for Detection, Quantification, and Analysis of Breast Cancer Biomarker Concentrations in Blood Serum
doi: 10.3390/bios15070447
Figure Lengend Snippet: ( A ) UV–Vis spectra showing the stepwise functionalization and bioconjugation of gold GNUs exhibiting an LSPR peak at 675 nm, PEG-functionalized GNUs, ADH-functionalized GNU-PEG with a peak at 220 nm, and mAb-conjugated GNU-PEG-ADH displaying a peak at 230 nm followed by a characteristic peak at 214 nm after the interaction. ( B ) Magnified view of ( A ) between 200 and 300 nm. ( C ) Magnification of ( B ) between 270 and 290 nm, where the mAb shows higher absorbance than HER-II, and the corresponding results for CA 15-3 from ( D – F ). Note the higher absorbance of CA 15-3 than the mAb.
Article Snippet: HER-II and CA15-3 monoclonal
Techniques:
Journal: Biosensors
Article Title: Dual-Oriented Targeted Nanostructured SERS Label-Free Immunosensor for Detection, Quantification, and Analysis of Breast Cancer Biomarker Concentrations in Blood Serum
doi: 10.3390/bios15070447
Figure Lengend Snippet: SERS signals of standard HER-II solution for before (mAb-conjugated) and after interaction at various concentrations.
Article Snippet: HER-II and CA15-3 monoclonal
Techniques:
Journal: Biosensors
Article Title: Dual-Oriented Targeted Nanostructured SERS Label-Free Immunosensor for Detection, Quantification, and Analysis of Breast Cancer Biomarker Concentrations in Blood Serum
doi: 10.3390/bios15070447
Figure Lengend Snippet: The trials of the SERS results of the functionalization, conjugation, and interaction steps for (GNU-mAb)-HER-II.
Article Snippet: HER-II and CA15-3 monoclonal
Techniques: Conjugation Assay
Journal: Biosensors
Article Title: Dual-Oriented Targeted Nanostructured SERS Label-Free Immunosensor for Detection, Quantification, and Analysis of Breast Cancer Biomarker Concentrations in Blood Serum
doi: 10.3390/bios15070447
Figure Lengend Snippet: SERS signals of standard CA15-3 solutions for before (mAb-conjugated) and after interaction at various concentrations.
Article Snippet: HER-II and CA15-3 monoclonal
Techniques:
Journal: Biosensors
Article Title: Dual-Oriented Targeted Nanostructured SERS Label-Free Immunosensor for Detection, Quantification, and Analysis of Breast Cancer Biomarker Concentrations in Blood Serum
doi: 10.3390/bios15070447
Figure Lengend Snippet: The trials of the SERS results of the functionalization, conjugation, and interaction steps for (GNU-mAb)-CA15-3.
Article Snippet: HER-II and CA15-3 monoclonal
Techniques: Conjugation Assay
Journal: Biosensors
Article Title: Dual-Oriented Targeted Nanostructured SERS Label-Free Immunosensor for Detection, Quantification, and Analysis of Breast Cancer Biomarker Concentrations in Blood Serum
doi: 10.3390/bios15070447
Figure Lengend Snippet: SERS signals of CA15-3 BCS samples for before (mAb-conjugated) and after interaction at various concentrations.
Article Snippet: HER-II and CA15-3 monoclonal
Techniques:
Journal: Biosensors
Article Title: Dual-Oriented Targeted Nanostructured SERS Label-Free Immunosensor for Detection, Quantification, and Analysis of Breast Cancer Biomarker Concentrations in Blood Serum
doi: 10.3390/bios15070447
Figure Lengend Snippet: PCA plots for various stages of interaction: ( A ) GNU-mAb + HER-II, ( B ) GNU-mAb + CA15-3, and ( C ) GNU-mAb+HBS.
Article Snippet: HER-II and CA15-3 monoclonal
Techniques:
Journal: JNCI Journal of the National Cancer Institute
Article Title: Quantitative Assessment of Effect of Preanalytic Cold Ischemic Time on Protein Expression in Breast Cancer Tissues
doi: 10.1093/jnci/djs438
Figure Lengend Snippet: Biomarkers used in the study*
Article Snippet: Vendor Biomarkers for companion diagnostic tests Estrogen receptor-alpha (ERα) Mouse 1D5/IgG1kappa M7047 DAKO Rabbit SP1/IgG RM-9101 Thermo Scientific Progesterone receptor (PgR) Mouse PgR636/IgG1kappa M3569 DAKO Rabbit PgRA/B (C89F7) 3153
Techniques: Diagnostic Assay, Modification, Purification