Journal: International journal of cancer

Article Title: Difference in mesothelin-binding ability of serum CA125 between patients with endometriosis and epithelial ovarian cancer.

doi: 10.1002/ijc.29185

Figure Lengend Snippet: Figure 1. A sandwich ELISA involving a recombinant mesothelin and anti-CA125 mAb (CA125meso assay). (a) A schematic diagram of the CA125meso assay. (b) Linearity study of the CA125meso assay. Serum specimens from patients with endometriosis (n 5 2) and EOC (n 5 2), and OVCAR-3 cells-produced CA125 were diluted to give the indicated concentrations of CA125 and then analyzed by means of the CA125meso assay. (c and d) The serum levels of CA125meso (c) and CA125 (d) were examined in patients with endometriosis (n 5 59) and EOC (n 5 36). The horizontal dotted lines represent the arbitrary cutoff values; 15 U/ml in (c) and 35 U/ml in (d). (e) ROC curves for the CA125 and CA125meso assays for discriminating endometriosis from EOC.

Article Snippet: For measurement of serum CA125meso, the wells of Maxisorp Immunoplates (Nunc, Roskilde, Denmark) were coated with a recombinant human mesothelin (R&D Systems, Minneapolis, MN).

Techniques: Sandwich ELISA, Recombinant, Produced

Journal: International journal of cancer

Article Title: Difference in mesothelin-binding ability of serum CA125 between patients with endometriosis and epithelial ovarian cancer.

doi: 10.1002/ijc.29185

Figure Lengend Snippet: Figure 2. Evaluation of the mesothelin-binding ability of CA125 (CA125meso/CA125). (a) Comparison of the CA125meso/CA125 levels between patients with endometriosis and EOC. (b) Relationship between the serum CA125 and CA125meso/CA125 levels in patients with EOC. (c) Compar- ison of the CA125meso/CA125 levels between premenopausal and postmenopausal patients with EOC. (d) Comparison of the CA125meso/CA125 levels according to the clinical stage of EOC. (e) Comparison of the CA125meso/CA125 levels according to the histological type of EOC. In (a) and (c)–(e), the horizontal bars represent the mean values. In (a), (d) and (e), the horizontal dotted line represents the arbitrary cutoff value (1.08).

Article Snippet: For measurement of serum CA125meso, the wells of Maxisorp Immunoplates (Nunc, Roskilde, Denmark) were coated with a recombinant human mesothelin (R&D Systems, Minneapolis, MN).

Techniques: Binding Assay, Comparison

Journal: International journal of cancer

Article Title: Difference in mesothelin-binding ability of serum CA125 between patients with endometriosis and epithelial ovarian cancer.

doi: 10.1002/ijc.29185

Figure Lengend Snippet: Figure 3. Effects of soluble mesothelin and MUC1 on the measurement of CA125meso. (a and d) The serum levels of mesothelin (a) and CA15- 3 antigen (d) were determined in patients with endometriosis (n 5 59 or 32) and EOC (n 5 36 or 30). (b and e) Relationship between the CA125meso/CA125 levels and the endogenous serum mesothelin (b) or CA15-3 (e) level. (c) OVCAR-3 cells-produced CA125 was diluted to 35 U/ml and then analyzed by means of the CA125meso assay in the presence of the indicated concentrations of the recombinant soluble human mesothelin (0–640 ng/ml). The data represent the means6 SE (n 5 4). In (a) and (d), the horizontal bars represent the mean values.

Article Snippet: For measurement of serum CA125meso, the wells of Maxisorp Immunoplates (Nunc, Roskilde, Denmark) were coated with a recombinant human mesothelin (R&D Systems, Minneapolis, MN).

Techniques: Produced, Recombinant

Journal: Journal for Immunotherapy of Cancer

Article Title: Chimeric antigen receptor T cells engineered to recognize the P329G-mutated Fc part of effector-silenced tumor antigen-targeting human IgG1 antibodies enable modular targeting of solid tumors

doi: 10.1136/jitc-2022-005054

Figure Lengend Snippet: IgG and soluble recombinant MSLN do not activate P329G-targeting CAR T cells. (A) IFN-γ ELISA after 96 hours of co-culture for one representative donor out of three independent donors. CAR T cells were cultivated with SUIT-2-MSLN cells and P329G LALA-mutated EGFR (EGFR-PG) or glycoengineered EGFR (EGFR-GE) binder in a concentration of 1.0 or 0.1 µg/mL in the absence or presence of IgG at a concentration of 1 mg/mL. SUIT-2-MSLN tumor cell killing by (B) P329G-targeting CAR (P329G-CAR) T cells and (C) CD16-ECD-CAR (CD16-CAR) T cells in different EGFR binder conditions measured over time through xCELLigence technology. (D) SUIT-2-MSLN tumor cell killing by P329G-CAR T cells and MSLN binder in absence or presence of soluble MSLN (100 ng/mL). (E) IFN-γ ELISA after 96 hours of co-culture for three independent donors (n=3). P329G-CAR T cells and MSLN binder (1.0 or 0.1 µg/mL) were cultivated with SUIT-2-MSLN in presence or absence of soluble MSLN (100 ng/mL). (F) IFN-γ ELISA after 24 hours of co-culture for three independent donors (n=3). P329G-CAR T cells and MSLN binder were cultivated in presence or absence of plate-bound MSLN (5 µg/mL) or soluble MSLN (0, 10, 100, or 1000 ng/mL). Each experiment was performed in triplicates. Subfigures (A–D) show data of one representative donor out of three independent experiments. Values in all graphs represent means±SEM (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). P values for xCELLigence data in (B–D) are shown for last time point. Only selected p values are shown.

Article Snippet: Flat bottom 96-well plates were coated overnight at 4°C with Fc-tagged recombinant human MSLN or recombinant HER2/ErbB2 (SinoBiological, China) in a concentration of 5 µg/mL diluted in 50 or 100 µL phosphate-buffered saline (PBS).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Concentration Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Chimeric antigen receptor T cells engineered to recognize the P329G-mutated Fc part of effector-silenced tumor antigen-targeting human IgG1 antibodies enable modular targeting of solid tumors

doi: 10.1136/jitc-2022-005054

Figure Lengend Snippet: P329G-targeting CAR T cells combined with the MSLN binder mediate in vitro and in vivo effector functions. (A) Schematic representation of P329G-targeting CAR (P329G-CAR), MSLN-targeting CAR (MSLN-CAR), and CD33-targeting CAR (CD33-CAR). (B) Density of MSLN surface expression on SUIT-2-MSLN and MSTO-MSLN-LUC cells. (C) MSTO-MSLN-LUC tumor cell killing by different P329G-CAR T cell conditions measured by luciferase-based assay after 64 hours of co-culture for three independent donors (n=3). (D) SUIT-2-MSLN, MSTO-MSLN-LUC and MCF-7 HER2 OE tumor cell killing by different T cell conditions measured over time through xCELLigence. (E) IFN-γ ELISA after 48 hours of co-culture with SUIT-2-MSLN, MSTO-MSLN-LUC and MCF-7 HER2 OE for one representative donor out of three independent donors. (F) Experimental layout for (G, H). MSLN binder was administered intraperitoneally (i.p.) twice a week. (G, H) MSTO-MSLN-LUC tumor growth curves of mice treated with a single intravenous injection of phosphate-buffered saline (PBS) or 10 7 CAR T cells (n=5 mice per group). (G, H) MSLN binder-treated mice received 20 µg or 5 µg per mouse. Each experiment was performed in triplicates. Subfigures (D, E) show data of one representative donor out of three independent experiments. Values in all graphs represent means±SEM (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). P values for xCELLigence data in (D) are shown for last time point. Only selected p values are shown.

Article Snippet: Flat bottom 96-well plates were coated overnight at 4°C with Fc-tagged recombinant human MSLN or recombinant HER2/ErbB2 (SinoBiological, China) in a concentration of 5 µg/mL diluted in 50 or 100 µL phosphate-buffered saline (PBS).

Techniques: In Vitro, In Vivo, Expressing, Luciferase, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Injection

Journal: Journal for Immunotherapy of Cancer

Article Title: Chimeric antigen receptor T cells engineered to recognize the P329G-mutated Fc part of effector-silenced tumor antigen-targeting human IgG1 antibodies enable modular targeting of solid tumors

doi: 10.1136/jitc-2022-005054

Figure Lengend Snippet: P329G-targeting CAR T cells mediated effector functions in a modular and reversible fashion. (A–C) IFN-γ ELISA after co-culture of P329G-targeting CAR (P329G-CAR), HER2-targeting CAR (HER2-CAR), MSLN-targeting CAR (MSLN-CAR) T cells or untransduced T cells (UT) with recombinant HER2 and different conditions of binders for three independent donors (n=3). (D–F) IFN-γ ELISA after co-culture of CAR T cells or UT with recombinant MSLN and different conditions of binders for three independent donors (n=3). (G) Experimental layout for (H). HER2 or MSLN binder (5 µg per mouse) were administered intraperitoneally (i.p.) twice a week. Bleeding and organ isolation was performed for mice still alive on day 28 after MSTO-MSLN-LUC tumor cell injection. (H) MSTO-MSLN-LUC tumor growth curves of survivor mice re-challenged with the tumor (n=4 mice per group) and treatment naïve mice (n=4–5 mice per group). (I) Human T cells and CAR T cells per beads ratio for blood, spleen, and tumor of survivor mice treated with P329G-CAR T cells and the binders. Each experiment of subfigures (A–F) was performed in triplicates. Values in all graphs represent means±SEM (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001). Only selected p values are shown.

Article Snippet: Flat bottom 96-well plates were coated overnight at 4°C with Fc-tagged recombinant human MSLN or recombinant HER2/ErbB2 (SinoBiological, China) in a concentration of 5 µg/mL diluted in 50 or 100 µL phosphate-buffered saline (PBS).

Techniques: Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Recombinant, Isolation, Injection

Journal: Scientific Reports

Article Title: Comparative analysis and process optimization for manufacturing CAR-T using the PiggyBac system derived from cryopreserved versus fresh PBMCs

doi: 10.1038/s41598-025-89686-7

Figure Lengend Snippet: The antibodies employed for staining and flow cytometric analysis of cell phenotypes.

Article Snippet: Biotin Human Mesothelin protein , Genscript Biotech, Nanjing, China , N/A.

Techniques: Staining