human mesothelin elisa kit  (R&D Systems)

Journal: Clinical and Experimental Medicine

Article Title: A proteomics approach to identify predictive blood biomarkers for pleural mesothelioma in prospective cohorts

doi: 10.1007/s10238-026-02058-x

Figure Lengend Snippet: Proteomics Workflow in EPIC and MoMar Cohorts. Left (EPIC): SWATH LC-MS/MS profiled 323 plasma proteins from 21 asbestos-exposed cases (PM within 5 years) and 21 matched controls. A GLM identified 12 DEPs ( p ≤ 0.05, fold change ≥ 1.3 ≤ 0.75); four were ELISA-validated. Calretinin and mesothelin benchmark ELISA assays were performed. Combined markers underwent ROC analysis Right (MoMar): Three DEPs from EPIC were quantified by ELISA alongside calretinin and mesothelin. ROC analysis of the combined protein panel assessed reproducibility and predictive accuracy in the pre-diagnosis cohort

Article Snippet: The measurements of serum calretinin and mesothelin were determined by ELISA assays in the EPIC samples using the Calretinin ELISA kit (DLD Diagnostika, Hamburg, Germany) and Human Mesothelin ELISA kit (R&D systems Inc.) as previously described [ ].

Techniques: Data-independent acquisition, Liquid Chromatography with Mass Spectroscopy, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Clinical and Experimental Medicine

Article Title: A proteomics approach to identify predictive blood biomarkers for pleural mesothelioma in prospective cohorts

doi: 10.1007/s10238-026-02058-x

Figure Lengend Snippet: ELISA validation of candidate DEPs in pre-diagnostic EPIC sera. Box plots display serum concentrations of B2M (ng/mL), C4 (µg/mL), TF (µg/mL), and DCD (µg/mL) in cases sampled 0–2 years and 2–5 years before PM diagnosis versus matched controls. B2M showed a borderline significant increase in the 0–2 year group (t-test and GLM p < 0.05); TF, C4, and DCD trended similarly but were not significant. Asterisks indicate nominal p < 0.05

Article Snippet: The measurements of serum calretinin and mesothelin were determined by ELISA assays in the EPIC samples using the Calretinin ELISA kit (DLD Diagnostika, Hamburg, Germany) and Human Mesothelin ELISA kit (R&D systems Inc.) as previously described [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Biomarker Discovery, Diagnostic Assay

Journal: Clinical and Experimental Medicine

Article Title: A proteomics approach to identify predictive blood biomarkers for pleural mesothelioma in prospective cohorts

doi: 10.1007/s10238-026-02058-x

Figure Lengend Snippet: ELISA replication of candidate DEPs in MoMar plasma. Box plots show concentrations of B2M (ng/mL), TF (µg/mL), and C4 (µg/mL) in pre-diagnostic cases versus asbestos-exposed non-cancer controls. No significant differences were observed for any marker

Article Snippet: The measurements of serum calretinin and mesothelin were determined by ELISA assays in the EPIC samples using the Calretinin ELISA kit (DLD Diagnostika, Hamburg, Germany) and Human Mesothelin ELISA kit (R&D systems Inc.) as previously described [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Diagnostic Assay, Marker

Journal: Clinical and Experimental Medicine

Article Title: A proteomics approach to identify predictive blood biomarkers for pleural mesothelioma in prospective cohorts

doi: 10.1007/s10238-026-02058-x

Figure Lengend Snippet: ELISA quantification of calretinin and mesothelin in pre-diagnostic EPIC and MoMar samples. ( A ) EPIC serum: calretinin (ng/mL) and mesothelin (nM) levels in cases sampled 0–2 years and 2–5 years before diagnosis versus controls. ( B ) MoMar plasma: calretinin and mesothelin levels in 0–2 years and 2–5 years pre-diagnostic cases compared to asbestos-exposed non-cancer controls. Calretinin was significantly elevated in MoMar cases at 0–2 years ( p < 0.01), whereas EPIC increases were not significant. Mesothelin levels reached significance only in MoMar ( p < 0.0001). Asterisks indicate significance (** p < 0.01; *** p < 0.0001)

Article Snippet: The measurements of serum calretinin and mesothelin were determined by ELISA assays in the EPIC samples using the Calretinin ELISA kit (DLD Diagnostika, Hamburg, Germany) and Human Mesothelin ELISA kit (R&D systems Inc.) as previously described [ ].

Techniques: Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Biomarker Discovery, Clinical Proteomics

Journal: Clinical and Experimental Medicine

Article Title: A proteomics approach to identify predictive blood biomarkers for pleural mesothelioma in prospective cohorts

doi: 10.1007/s10238-026-02058-x

Figure Lengend Snippet: Receiver operating characteristics (ROC) analysis of combined biomarker panels in pre-diagnostic PM cases. ( A ) EPIC cohort: ROC curves for models using B2M alone (mod1, AUC = 0.71), B2M + C4 + TF (mod2, AUC = 0.81), Mesothelin + Calretinin (mod3, AUC = 0.65), and all five markers (mod4, AUC = 0.88; p-value = 0.17 vs. mod1 by DeLong’s test). ( B ) MoMar cohort: ROC curves for the same models (mod1 AUC = 0.55), mod2 (AUC = 0.75), mod3 (AUC = 0.84), and mod4 (AUC = 0.91; p = 0.001 vs. mod1)

Article Snippet: The measurements of serum calretinin and mesothelin were determined by ELISA assays in the EPIC samples using the Calretinin ELISA kit (DLD Diagnostika, Hamburg, Germany) and Human Mesothelin ELISA kit (R&D systems Inc.) as previously described [ ].

Techniques: Biomarker Discovery, Diagnostic Assay

Journal: The FASEB Journal

Article Title: Excess shed mesothelin disrupts pancreatic cancer cell clustering to impair peritoneal colonization

doi: 10.1096/fj.202400446R

Figure Lengend Snippet: Assessing the activity of MPF versus membrane‐bound mature MSLN. KLM1 MSLN KO cells were stably transfected with empty vector (KO+vec), MSLNf, or MPFf. (A) KLM1 derivative cells were assessed for membrane‐bound MSLN expression by flow cytometry. (B) Growth rate of the cell lines on tissue culture plastic was measured. There was no significant difference between the groups. (C–F) Cell lines were injected IP into nude mice and allowed to grow for ~6 weeks. (C) Total burden of peritoneal tumor was measured, *** p < .001. (D, E) MSLN and MPF concentration in tumor lysate was measured by ELISA assay. (F) Tumor burden of co‐injected +MSLNf and +MPFf cells was assessed, * p < .05; ns, not significant.

Article Snippet: All serum/media concentrations were measured using Quantikine ELISA Kits (R&D Systems, for MSLN #DMSLN0, IL‐1α #DLA50, LIF #DLF00B).

Techniques: Activity Assay, Membrane, Stable Transfection, Transfection, Plasmid Preparation, Expressing, Flow Cytometry, Injection, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: The FASEB Journal

Article Title: Excess shed mesothelin disrupts pancreatic cancer cell clustering to impair peritoneal colonization

doi: 10.1096/fj.202400446R

Figure Lengend Snippet: Assessing the activity of secreted‐only MSLN in KO cells. (A) Schema depicting truncation mutant (Δ591) to remove the GPI anchoring site and prevent membrane association of mature MSLN. KLM1 MSLN KO cells were stably transduced with WT and Y318A (Mu) Δ591 expression vectors and single cell clones were isolated. (B) Immunoblot of Δ591 clones to assess for MSLN expression. (C) Conditioned medium of Δ591 clones plated at equal density was assayed for MSLN concentration by ELISA. (D) Representative experiment showing growth rate of the KO+ Δ591 cell lines on tissue culture plastic as measured by counting cell number in triplicate wells. (E) Cells were suspended in soft agar and colonies were counted after ~3 weeks. Figure depicts raw data of triplicate wells (marker) in each experiment (bar). ** p < .01 or *** p < .001 indicate statistically significant difference as compared to parent for at least 2 of 3 experiments (in same direction of change). (F, G) Cell lines were injected IP into nude mice and allowed to grow for ~6 weeks. (F) Tumors were lysed and MSLN expression was assayed by immunoblot. (G) Total burden of peritoneal tumor dissected from the mouse abdominal cavity, * p < .05. Each point represents one animal. Results were confirmed by repeat using other Δ591WT and Δ591Mu clones. (H) MUC‐16 surface expression was assessed by flow cytometry. Left— Representative tracing. Right— Summary of geometric means in relation to Parent over multiple experiments.

Article Snippet: All serum/media concentrations were measured using Quantikine ELISA Kits (R&D Systems, for MSLN #DMSLN0, IL‐1α #DLA50, LIF #DLF00B).

Techniques: Activity Assay, Mutagenesis, Membrane, Stable Transfection, Transduction, Expressing, Clone Assay, Isolation, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay, Marker, Injection, Flow Cytometry

Journal: The FASEB Journal

Article Title: Excess shed mesothelin disrupts pancreatic cancer cell clustering to impair peritoneal colonization

doi: 10.1096/fj.202400446R

Figure Lengend Snippet: Overexpression of sMSLN inhibits the pro‐tumorigenic activity of cells expressing membrane‐bound MSLN. (A) Proposed model showing how sMSLN might block cell–cell association by interfering with interactions between MUC‐16 and membrane‐bound MSLN. (B–H) KLM1 cells were stably transduced with Δ591 MSLN expression vectors and pooled cells were used for experiments. (B) Expression of MSLN in conditioned medium as detected by ELISA. (C) Representative experiment showing growth rate of the cell lines on tissue culture plastic as measured by counting cell number in triplicate wells for each time point. No significant difference in growth was observed. (D–F) Cell lines were injected IP into nude mice and allowed to grow for ~6 weeks, * p < .05; *** p < .001; **** p < .0001; ns, not significant. (D) Total burden of peritoneal tumor dissected from the mouse abdominal cavity. Each point represents one animal. (E) Tumors were lysed and MSLN expression was assayed by immunoblot. Each lane is lysate from one mouse tumor. (F) Serum MSLN expression was assayed by ELISA. (G, H) Indicated cells were tagged with CellTracker dye and visualized by fluorescence microscopy. Representative images of multiple replicates with four to eight fields imaged per replicate. (G) Labeled cells were plated at equal density onto low adherence plates for 24 h before visualization. (H) Labeled cells were injected IP into nude mice. Mice were euthanized 4 h later and peritoneal lavage fluid was collected for visualization.

Article Snippet: All serum/media concentrations were measured using Quantikine ELISA Kits (R&D Systems, for MSLN #DMSLN0, IL‐1α #DLA50, LIF #DLF00B).

Techniques: Over Expression, Activity Assay, Expressing, Membrane, Blocking Assay, Stable Transfection, Transduction, Enzyme-linked Immunosorbent Assay, Injection, Western Blot, Fluorescence, Microscopy, Labeling

Journal: The FASEB Journal

Article Title: Excess shed mesothelin disrupts pancreatic cancer cell clustering to impair peritoneal colonization

doi: 10.1096/fj.202400446R

Figure Lengend Snippet: Excess of sMSLN prevents cell clustering. (A) Schema showing method used to produce MSLN‐containing CM. Mock cells underwent transient transfection with Cas9 vector like MSLN KO cells, but no gRNA was included, so they retain endogenous MSLN expression (MSLN+). Transduction of KO cells with full‐length MSLN WT or Y318A expression construct produces the KO+WT and KO+Mu cell lines which overexpress MSLN (MSLN++). (B) ELISA to assess MSLN concentration of CM from equal numbers of cells plated for each type. (C) Cells were tagged with CellTracker dye then plated at equal density onto low adherence plates for 24 h before visualization by fluorescence microscopy. Representative images of multiple replicates with 4–8 fields imaged per replicate. (D) Schema for treatment of KLM1 and T3M4 KO cells with CM prior to RNA harvest. (E) PCA plots following RNA deep‐sequencing of CM‐treated KO cells. (F) Quantitation and directionality of GSEA pathway changes in KO cells treated with KO CM (negative control) as compared to MSLN‐containing CMs.

Article Snippet: All serum/media concentrations were measured using Quantikine ELISA Kits (R&D Systems, for MSLN #DMSLN0, IL‐1α #DLA50, LIF #DLF00B).

Techniques: Transfection, Plasmid Preparation, Expressing, Transduction, Construct, Enzyme-linked Immunosorbent Assay, Concentration Assay, Fluorescence, Microscopy, Sequencing, Quantitation Assay, Negative Control

Journal: The FASEB Journal

Article Title: Excess shed mesothelin disrupts pancreatic cancer cell clustering to impair peritoneal colonization

doi: 10.1096/fj.202400446R

Figure Lengend Snippet: Soluble MSLN exposure results in release of IL‐1α. (A, B) Venn diagrams outlining GSEA pathway changes that occur when MSLN KO cells are exposed to sMSLN‐containing CM as compared with exposure to CM from MSLN KO cells. Pathways with statistically significant differential expression are listed. Those in bold were significantly changed in both KLM1 and T3M4. (C, D) KO cells were treated with stock CM isolated from KO, Mock, +WT or +Mu cells for 0 or 4 h, ** p < .01; **** p < .0001; ns, not significant. The concentration of IL‐1α (C) or LIF (D) in CM was then measured by ELISA.

Article Snippet: All serum/media concentrations were measured using Quantikine ELISA Kits (R&D Systems, for MSLN #DMSLN0, IL‐1α #DLA50, LIF #DLF00B).

Techniques: Quantitative Proteomics, Isolation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Hyaluronan and N-ERC/Mesothelin as Key Biomarkers in a Specific Two-Step Model to Predict Pleural Malignant Mesothelioma

doi: 10.1371/journal.pone.0072030

Figure Lengend Snippet: Levels of hyaluronan, N-ERC/mesothelin, C-ERC/mesothelin, osteopontin, syndecan-2, syndecan-1, and thioredoxin in patients with malignant pleural mesothelioma, other malignant pleural disease, or benign effusions. Values of 0 (i.e. below the detection limit) are tabulated for each group under the respective graph, as they cannot be shown on a logarithmic scale. Dotted line represents cut-off values. Horizontal lines represent medians. N tot /biomarker<190 indicates the exclusion of patients from individual analyses due to insufficient material (e.g. thioredoxin, n = 186).

Article Snippet: ELISA kits for human N-ERC/mesothelin (code no. 99666/7–16 assay), osteopontin (code no. 27158) and thioredoxin (code no. 27417) were all purchased from Immuno-Biological Laboratories Co., Ltd., Japan.

Techniques: Biomarker Assay

Journal: PLoS ONE

Article Title: Hyaluronan and N-ERC/Mesothelin as Key Biomarkers in a Specific Two-Step Model to Predict Pleural Malignant Mesothelioma

doi: 10.1371/journal.pone.0072030

Figure Lengend Snippet: Odds ratios for biomarkers in the model generation dataset.

Article Snippet: ELISA kits for human N-ERC/mesothelin (code no. 99666/7–16 assay), osteopontin (code no. 27158) and thioredoxin (code no. 27417) were all purchased from Immuno-Biological Laboratories Co., Ltd., Japan.

Techniques:

Journal: PLoS ONE

Article Title: Hyaluronan and N-ERC/Mesothelin as Key Biomarkers in a Specific Two-Step Model to Predict Pleural Malignant Mesothelioma

doi: 10.1371/journal.pone.0072030

Figure Lengend Snippet: A) ROC curves showing sensitivity and specificity for individual biomarkers. B) Predicted risk values from two-step model based on hyaluronan and N-ERC/mesothelin. Cases with a predicted risk>0.9 were considered positive (above shaded area). C) ROC curve for the two-step model.

Article Snippet: ELISA kits for human N-ERC/mesothelin (code no. 99666/7–16 assay), osteopontin (code no. 27158) and thioredoxin (code no. 27417) were all purchased from Immuno-Biological Laboratories Co., Ltd., Japan.

Techniques:

Journal: PLoS ONE

Article Title: Hyaluronan and N-ERC/Mesothelin as Key Biomarkers in a Specific Two-Step Model to Predict Pleural Malignant Mesothelioma

doi: 10.1371/journal.pone.0072030

Figure Lengend Snippet: A) and B) Levels of hyaluronan and N-ERC/mesothelin, respectively, in the validation dataset. The dotted line represents cut-off values. C) Predicted risk values from the two-step predictive model. Cases with predicted risks>0.9 were considered positive (above shaded area). D) ROC curves generated from N-ERC/mesothelin (solid black line) and hyaluronan (dotted black line) as single markers or combined in the two-step model (dotted grey line).

Article Snippet: ELISA kits for human N-ERC/mesothelin (code no. 99666/7–16 assay), osteopontin (code no. 27158) and thioredoxin (code no. 27417) were all purchased from Immuno-Biological Laboratories Co., Ltd., Japan.

Techniques: Generated

Journal: PLoS ONE

Article Title: Hyaluronan and N-ERC/Mesothelin as Key Biomarkers in a Specific Two-Step Model to Predict Pleural Malignant Mesothelioma

doi: 10.1371/journal.pone.0072030

Figure Lengend Snippet: A) Schematic presentation of the two-step model and its performance on the model generation dataset. B) Schematic presentation of the two-step model and its performance on the validation dataset. In both A) and B) , after the logistic regression a predicted risk value>0.9 indicates additional mesothelioma cases compared to hyaluronan or N-ERC/mesothelin alone. C) and D) Calibration plots showing the agreement between observed outcomes (y-axis) and predictions (x-axis) in the model generation dataset and validation dataset respectively.

Article Snippet: ELISA kits for human N-ERC/mesothelin (code no. 99666/7–16 assay), osteopontin (code no. 27158) and thioredoxin (code no. 27417) were all purchased from Immuno-Biological Laboratories Co., Ltd., Japan.

Techniques:

Journal: PLoS ONE

Article Title: Hyaluronan and N-ERC/Mesothelin as Key Biomarkers in a Specific Two-Step Model to Predict Pleural Malignant Mesothelioma

doi: 10.1371/journal.pone.0072030

Figure Lengend Snippet: Box plots of predicted probabilities using hyaluronan, N-ERC/mesothelin or the two-step model on the model generation dataset and validation dataset. The mean is denoted by a “+”, whiskers indicate the 5 th and 95 th percentiles, while dots are outliers. Grey dotted lines represent the discrimination slope (DS) which is equivalent to the integrated discrimination index. The differences in the discrimination slopes correspond to the integrated discrimination index , . As an example, the highest possible discrimination slope would be 1 (100%) and comparing N-ERC/mesothelin with the two-step model in the model generation dataset shows that the integrated discrimination index increases with 20% when applying the two-step model (DS Model 0.84 minus DS N-ERC 0.64).

Article Snippet: ELISA kits for human N-ERC/mesothelin (code no. 99666/7–16 assay), osteopontin (code no. 27158) and thioredoxin (code no. 27417) were all purchased from Immuno-Biological Laboratories Co., Ltd., Japan.

Techniques: