Journal: PLOS ONE

Article Title: NAV-001, a high-efficacy antibody-drug conjugate targeting mesothelin with improved delivery of a potent payload by counteracting MUC16/CA125 inhibitory effects

doi: 10.1371/journal.pone.0285161

Figure Lengend Snippet: Antibodies were screened via ELISA for binding to immobilized MSLN and MUC16/CA125 proteins. Human serum albumin (HSA) was used as negative control. Panel A shows that the antibody referred to herein as NAV-001 was the only antibody from the group that did not bind to MUC16/CA125 protein (P = 0.000025). To determine if MUC16/CA125 perturbs anti-MSLN binding to MSLN protein, immobilized MSLN was probed with each antibody with or without MUC16/CA125 (CA). As shown in panel B, MUC16/CA125 had no impact on antibodies binding to MSLN. All data represent a minimum of triplicate experiments.

Article Snippet: To confirm MSLN and MUC16/CA125 expression in PDFs, we employed IHC using a rabbit anti-MSLN antibody that binds the same epitope as NAV-001 and a commercial rabbit anti-MUC16/CA125 antibody (Novus), respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Negative Control

Journal: PLOS ONE

Article Title: NAV-001, a high-efficacy antibody-drug conjugate targeting mesothelin with improved delivery of a potent payload by counteracting MUC16/CA125 inhibitory effects

doi: 10.1371/journal.pone.0285161

Figure Lengend Snippet: Antibodies Ab-1, Ab-2 and NAV-001 were converted into SN-38 ADCs and tested for cytotoxicity using the isogenic MSLN-expressing OVCAR-3 and OV-KD cells, the latter which lacks MUC16/CA125 expression. As shown in Panel A, Ab-1 and Ab-2 had reduced OVCAR-3 target cell (CA125 + ) killing as compared to NAV-001 (P < 0.00011). All three ADCs killed the isogenic OV-KD cells (CA125 - ) at a similar magnitude confirming the negative effect of MUC16/CA125 on ADCs containing antibodies that bind it. Shown is the differential killing of each ADC when cultures were treated with 50 ng/mL of each ADC. Panel B is a schematic representation of the potential mechanism by which MUC16/CA125 suppresses ADC killing through antibody binding and reduced internalization. All data represent a minimum of triplicate experiments.

Article Snippet: To confirm MSLN and MUC16/CA125 expression in PDFs, we employed IHC using a rabbit anti-MSLN antibody that binds the same epitope as NAV-001 and a commercial rabbit anti-MUC16/CA125 antibody (Novus), respectively.

Techniques: Expressing, Binding Assay

Journal: PLOS ONE

Article Title: NAV-001, a high-efficacy antibody-drug conjugate targeting mesothelin with improved delivery of a potent payload by counteracting MUC16/CA125 inhibitory effects

doi: 10.1371/journal.pone.0285161

Figure Lengend Snippet: Antibody 2 (Ab-2) and NAV-001 were converted into ADCs by linking the topoisomerase II cytotoxin PNU-159682 and both were tested for cytotoxicity against the isogenic MSLN-expressing OVCAR-3 (panel A and C) and OV-KD (panel B and D) cell lines. As shown in Panels A and B, a similar effect of target cell killing was observed as the SN-38 formatted ADCs shown in , whereby the MUC16/CA125 binding Ab-2-PNU had reduced killing in OVCAR-3 (A) as compared to NAV-001-PNU(P < 0.0033), while similar killing was observed for both against the isogenic non-MUC16/CA125-expressing OV-KD cells (B). Antibodies were tested for cellular uptake using the same cell pairs and as shown in Panels C (OVCAR-3) and D (OV-KD), Ab-2 has reduced internalization as compared to NAV-001 in OVCAR-3 cells (P < 0.00008) but similar internalization kinetics in the OV-KD cells. All data represent a minimum of triplicate experiments.

Article Snippet: To confirm MSLN and MUC16/CA125 expression in PDFs, we employed IHC using a rabbit anti-MSLN antibody that binds the same epitope as NAV-001 and a commercial rabbit anti-MUC16/CA125 antibody (Novus), respectively.

Techniques: Expressing, Binding Assay

Journal: PLOS ONE

Article Title: NAV-001, a high-efficacy antibody-drug conjugate targeting mesothelin with improved delivery of a potent payload by counteracting MUC16/CA125 inhibitory effects

doi: 10.1371/journal.pone.0285161

Figure Lengend Snippet: Panel 6A, PDFs used for each study were confirmed for MSLN and MUC16/CA125 expression by IHC analyses. As shown, all tumors had robust homogeneous MSLN staining, while the mesothelioma and TNBC tumors also expressed MUC16/CA125. These tumors served as models to confirm NAV-001-PNU in vivo efficacy against MUC16/CA125 expressing tumors. Panel B, tumor growth and tolerability of NAV-001 in PDX models. As shown, single-dose of NAV-001 at 0.25 or 0.75 mg/kg showed significant anti-tumor efficacy (P < 0.005), and long-term regression after 0.75 mg/kg single-dose regimen. Triangles represent day of dosing post randomization. Each cohort of each study employed 6 mice.

Article Snippet: To confirm MSLN and MUC16/CA125 expression in PDFs, we employed IHC using a rabbit anti-MSLN antibody that binds the same epitope as NAV-001 and a commercial rabbit anti-MUC16/CA125 antibody (Novus), respectively.

Techniques: Expressing, Staining, In Vivo

Journal: Journal of Cancer

Article Title: Tranexamic acid reduces endometrial cancer effects through the production of angiostatin

doi: 10.7150/jca.68169

Figure Lengend Snippet: Effects of TA treatment on plasminogen, plasmin and tPA levels. Thirty weeks after the start of the study, by western blotting, the expression of plasminogen (A) , plasmin (B) and tPA (C) in uterus on MNU- and estradiol-induced endometrial cancer model mice were measured. By ELISA kit, the levels of plasminogen (D) , plasmin (E) and tPA (F) in plasma on MNU- and estradiol-induced endometrial cancer model mice were measured. Values are expressed as the mean ± standard deviation (SD) derived from 10 animals. * p < 0.05. TA: tranexamic acid, tPA: tissue plasminogen activator, MNU: N -methyl- N -nitrosourea.

Article Snippet: The plasma levels of carbohydrate antigen 125 (CA125), interleukin (IL)-6, tumor necrosis factor (TNF-α), plasminogen, plasmin, and tPA were determined using commercial ELISA kits (CA125: Bioassay Technology Laboratory, Shanghai, China; Il-6: Proteintech, Rosemont, IL, USA; TNF-α: R&D Systems, Minneapolis, MN, USA; plasminogen and plasmin: LSBio, Seattle, WA, USA; tPA: Abcam, Cambridge, UK) according to the respective manufacturers' instructions.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Standard Deviation, Derivative Assay