anti cd30  (R&D Systems)

Journal: Cancer Chemotherapy and Pharmacology

Article Title: The combination of brentuximab vedotin and chidamide synergistically suppresses the proliferation of T-cell lymphoma cells through the enhancement of apoptosis

doi: 10.1007/s00280-023-04609-5

Figure Lengend Snippet: HH cells abundantly express CD30 and the viability is reduced after BV treatment. a Cell viabilities following treatment with BV (0.0068–68 nM) for 72 h. b Flow cytometry histograms. Cells were sorted following staining with PE-conjugated anti-CD30 antibodies (orange), with isotype-matched control antibodies (blue), or without antibodies (red). BV brentuximab vedotin, PE phycoerythrin

Article Snippet: Cells were collected and incubated for 30 min at 4 °C with phycoerythrin (PE)-labeled mouse anti-human CD30 antibodies (#130-098-686, Miltenyi Biotec, Bergisch Gladbach, Germany), with isotype-matched control antibodies (#130-092-213, Miltenyi Biotec), or without antibodies.

Techniques: Flow Cytometry, Staining, Control

Journal: Aesthetic surgery journal

Article Title: IL-9 Is a Biomarker of BIA-ALCL Detected Rapidly by Lateral Flow Assay.

doi: 10.1093/asj/sjae137

Figure Lengend Snippet: Figure 3. CD30 lateral flow assay. (A) CD30 standards were diluted from 0 to 10 ng/mL in a benign seroma. (B) Correlation of CD30 concentration with TL/CL ratio (r2 = 0.72, y = 0.05x + 0.7, P = .02). (C) Comparison of CD30 in benign and ALCL seromas (benign, 0.198 ± 0.036 [n = 15] vs ALCL, 0.779 ± 0.083 [n = 11]; P < .0001, t-test). CL, control line; TL, test line.

Article Snippet: Similarly, CD30 LFA strips were made by striping mouse anti-CD30 antibody (R&D Systems, catalog number MAB2291).

Techniques: Lateral Flow Assay, Concentration Assay, Comparison, Control

Journal: Molecular Therapy Oncolytics

Article Title: Nanobody-derived bispecific CAR-T cell therapy enhances the anti-tumor efficacy of T cell lymphoma treatment

doi: 10.1016/j.omto.2023.07.007

Figure Lengend Snippet: Construction of the Nb phage library and screening of anti-CD30 Nbs and anti-CD5 Nbs (A) Schematic of construction of the Nb phage library and screening of Nbs. (B) The variable domain of heavy chain of heavy-chain antibody (VHH) genes were obtained by two-step PCR, and the library size was measured by counting the colonies after serial dilution (M, DNA marker; 1, PCR product). (C) Monoclonal phage ELISA. 24 positive phages from the three rounds of panning were tested for binding affinity by ELISA with CD30- and CD5-coated plates. (D) Flow cytometry screening of monoclonal phages capable of binding to cells. Karpas-299 cells were used as target cells to validate positive monoclonal phages by flow cytometry. Negative control (NC) phages that did not express Nbs were used as a negative control.

Article Snippet: For target antigens staining, the following antibodies were used: mouse anti-human PE-CD30 (BD Biosciences, 550041) and mouse anti-human Brilliant Violet 421-CD5 (BD Biosciences, 562646).

Techniques: Serial Dilution, Marker, Enzyme-linked Immunosorbent Assay, Binding Assay, Flow Cytometry, Negative Control

Journal: Molecular Therapy Oncolytics

Article Title: Nanobody-derived bispecific CAR-T cell therapy enhances the anti-tumor efficacy of T cell lymphoma treatment

doi: 10.1016/j.omto.2023.07.007

Figure Lengend Snippet: In vitro binding ability validation of the screened Nbs (A) The biding affinity of Nb-Fc to CD30 or CD5 was measured by ELISA. (B) Three anti-CD30 Nbs and four anti-CD5 Nbs were selected, and we performed ELISA at the indicated dilution ratios by incubation with the corresponding antigens. Fc served as negative control. (C) Detection of the binding affinity of the Nb-Fc or scFv-Fc antibodies to CD5 or CD30, respectively, by ELISA. (D) Single-target NbCD30 or NbCD5-CAR-T cells were co-cultured with Karpas-299 cells (CD30 + CD5 + ) at different effector-to-target ratios for 24 h, and the specific cytotoxicity was determined by Lactic Dehydrogenase (LDH) assay. ∗∗∗∗p < 0.0001 for one-way ANOVA with multiple-comparisons test. All data are mean ± standard error.

Article Snippet: For target antigens staining, the following antibodies were used: mouse anti-human PE-CD30 (BD Biosciences, 550041) and mouse anti-human Brilliant Violet 421-CD5 (BD Biosciences, 562646).

Techniques: In Vitro, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation, Negative Control, Cell Culture, Lactate Dehydrogenase Assay

Journal: Molecular Therapy Oncolytics

Article Title: Nanobody-derived bispecific CAR-T cell therapy enhances the anti-tumor efficacy of T cell lymphoma treatment

doi: 10.1016/j.omto.2023.07.007

Figure Lengend Snippet: The binding ability was predicted by molecular docking and verified by SPR experiments (A) Molecular docking prediction model of anti-CD30/CD5 Nbs or scFv to corresponding antigens. The light blue structure indicates the antigen, the dark blue structure indicates the antibody, the interface area (red) indicates the region of the two proteins in contact with each other, and ΔG indicates the free energy of binding. (B) SPR experiments were performed to measure the binding affinity of Nb-Fc or scFv-Fc on its corresponding antigen.

Article Snippet: For target antigens staining, the following antibodies were used: mouse anti-human PE-CD30 (BD Biosciences, 550041) and mouse anti-human Brilliant Violet 421-CD5 (BD Biosciences, 562646).

Techniques: Binding Assay

Journal: Molecular Therapy Oncolytics

Article Title: Nanobody-derived bispecific CAR-T cell therapy enhances the anti-tumor efficacy of T cell lymphoma treatment

doi: 10.1016/j.omto.2023.07.007

Figure Lengend Snippet: Construction of bispecific Nb/scFv-derived CAR-T cells to verify in vitro function (A) Schematic of CAR structure and single- or bispecific scFv-CAR. (B) Schematic of single NbCD30/NbCD5-CAR or bispecific NbCD30-CD5-CAR. (C) Single-or bispecific scFv-CAR-T cells were co-cultured with Karpas-299 cells (CD30 + CD5 + ) or SupT1 cells (CD30 + CD5 − ) at the indicated E:T ratios for 24 h, and the specific cytotoxicity was determined by LDH assay. The data shown are representative results of three independent replication experiments. (D) Single-or bispecific Nb-CAR-T cells were co-cultured with Karpas-299 cells (CD30 + CD5 + ) or SupT1 cells (CD30 + CD5 − ) at different E:T ratios for 24 h, and the specific cytotoxicity was determined by LDH assay. The data shown are representative results of three independent replication experiments. (E) Bispecific Nb-CAR-T and scFv-CAR-T cells were co-cultured with Karpas-299 cells (CD30 + CD5 + ) or Raji cells (CD30 − CD5 − ) at different E:T ratios for 24 h, and the specific cytotoxicity was determined by LDH assay. (F) NbCD30/CD5-CAR-T cells or mock T cells (untransduced T cells) were incubated with tumor target cells (Karpas-299) for 24 h. The levels of IFN-γ, TNF-α, IL-2, and granzyme B in culture supernatants were measured by ELISA. (G) CAR-T cells or control T cells were incubated with tumor target cells (Karpas-299) at a 1:1 ratio for 24 h. Secretion of IFN-γ was measured by ELISpot assay. All data are representative of three independent replication experiments. All data are mean ± standard error; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 for one-way ANOVA.

Article Snippet: For target antigens staining, the following antibodies were used: mouse anti-human PE-CD30 (BD Biosciences, 550041) and mouse anti-human Brilliant Violet 421-CD5 (BD Biosciences, 562646).

Techniques: Derivative Assay, In Vitro, Cell Culture, Lactate Dehydrogenase Assay, Incubation, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

Journal: Molecular Therapy Oncolytics

Article Title: Nanobody-derived bispecific CAR-T cell therapy enhances the anti-tumor efficacy of T cell lymphoma treatment

doi: 10.1016/j.omto.2023.07.007

Figure Lengend Snippet: Bispecific CD30-CD5-CAR-T cells are more effective for tumor growth suppression (A and B) 6 × 10 6 Karpas-299 cells were inoculated subcutaneously into NCG mice. The tumor reached 100 mm 3 after 7–10 days. Various T cells were infused via tail vein injection. Tumor engraftment was monitored every 2–3 days (1–2 μg of human IL-2 per mouse was injected at a frequency of 2–3 days). s.c., subcutaneous; i.v., intravenous; i.p., intraperitoneal. Tumor volumes were monitored after injection of CAR-T cells (scale bar, 10 mm). (C) The tumor mass was quantified. All data are mean ± standard error; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 for one-way ANOVA.

Article Snippet: For target antigens staining, the following antibodies were used: mouse anti-human PE-CD30 (BD Biosciences, 550041) and mouse anti-human Brilliant Violet 421-CD5 (BD Biosciences, 562646).

Techniques: Injection

Journal: Molecular Therapy Oncolytics

Article Title: Nanobody-derived bispecific CAR-T cell therapy enhances the anti-tumor efficacy of T cell lymphoma treatment

doi: 10.1016/j.omto.2023.07.007

Figure Lengend Snippet: Nb-derived CD30 and CD5 bispecific CAR-T cells enhanced anti-tumor potency in vivo (A) Fluorescence-activated cell sorting (FACS) analysis shows the proportion of tumor-infiltrating CAR-T cells and cytokine production capacity. (B) Immunofluorescence staining of tumor tissue sections. The tumor tissue sections were analyzed for immunofluorescence by staining with anti-human CD8 (red), anti-human IFN-γ (green), and anti-human granzyme B (pink) antibodies, and the nuclei were stained with DAPI (blue). The statistical plot shows the mean fluorescence intensity statistics for CD8, IFN-γ, and granzyme B expressed as the mean ± SD from three randomly selected fields of thin tumor sections. Scale bar, 20 μm. (C) Survival curve of mice after tumor inoculation and then injection of the indicated CAR-T cells. ∗p < 0.05, ∗∗p < 0.01, log rank Mantel-Cox test. All data are mean ± standard error; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, one-way ANOVA.

Article Snippet: For target antigens staining, the following antibodies were used: mouse anti-human PE-CD30 (BD Biosciences, 550041) and mouse anti-human Brilliant Violet 421-CD5 (BD Biosciences, 562646).

Techniques: Derivative Assay, In Vivo, Fluorescence, FACS, Immunofluorescence, Staining, Injection