Journal: bioRxiv

Article Title: A MET-Targeted Variable New Antigen Receptor (VNAR) Theranostic for Non-Small Cell Lung Cancer

doi: 10.64898/2026.01.30.702875

Figure Lengend Snippet: ( A ) Designation of sites used for subcutaneous (s.c.) and intravenous (i.v.) delivery of immunogens and blood collection. ( B ) SDS-PAGE gel with Coomassie staining of extracellular domain of the MET protein with a See Blue ladder (left), the MET protein (center) and the MET protein under reducing conditions (right). ( C ) Diagram of the time course, injections, adjuvants, and blood collections throughout the MET immunization program. ( D ) Biolayer Interferometry (BLI) sensorgram from a representative experiment demonstrating the mobilization of an anti-MET immune response after MET immunization. Diluted plasma samples were collected over time and screened against biosensors loaded with MET to detect the presence of MET-binding antibodies. ( E ) Venn diagram of sequence overlap between NGS datasets derived from sequencing the MET-immunized VNAR phagemid library and a library constructed for an unrelated immunogen. ( F ) A column scatter of the number of amino acids in the CDR3 of VNARs by subtype, overlaid with box plots to illustrate the spread of the data.

Article Snippet: MET copy number variation was determined by predesigned TaqMan Copy Number Assay (Life Technologies, Hs04993403_cn; FAM-dye labeled) and TaqMan Copy Number Reference Assay as RNaseP (Life Technologies, #4403328, VIC-dye labeled). qPCR was performed using QuantStudio3 Real-Time PCR System (Applied Biosystems).

Techniques: SDS Page, Staining, Clinical Proteomics, Binding Assay, Sequencing, Derivative Assay, Construct

Journal: bioRxiv

Article Title: A MET-Targeted Variable New Antigen Receptor (VNAR) Theranostic for Non-Small Cell Lung Cancer

doi: 10.64898/2026.01.30.702875

Figure Lengend Snippet: ( A ) Dilution ELISA of promising VNAR clones demonstrate saturable binding to MET. Eleven unpurified VNARs were serially diluted and added to plates coated with MET protein. ( B ) BLI sensorgram of sensors loaded with a variety of proteins, each tested against a standard concentration of vMET1-Fc. ( C ) BLI sensorgrams of sensors loaded with human MET exposed to serially diluted vMET1 monomer or ( D ) vMET1-Fc, followed by dissociation in assay buffer. Dissociation constants (K D ) are listed for each assay.

Article Snippet: MET copy number variation was determined by predesigned TaqMan Copy Number Assay (Life Technologies, Hs04993403_cn; FAM-dye labeled) and TaqMan Copy Number Reference Assay as RNaseP (Life Technologies, #4403328, VIC-dye labeled). qPCR was performed using QuantStudio3 Real-Time PCR System (Applied Biosystems).

Techniques: Enzyme-linked Immunosorbent Assay, Clone Assay, Binding Assay, Concentration Assay

Journal: bioRxiv

Article Title: A MET-Targeted Variable New Antigen Receptor (VNAR) Theranostic for Non-Small Cell Lung Cancer

doi: 10.64898/2026.01.30.702875

Figure Lengend Snippet: ( A ) Heat map detailing in order top to bottom: site of origin and presence of oncogene, phosphorylated MET (p-MET) with respect to T-47D, MET Copy Number (CN), total MET protein expression with respect to T-47D, MET RNA, and MET receptor density. Numerical color spectrum is log-2 scaled. Black bar graph on top demonstrates vMET1-Fc binding quantified via flow cytometry normalized to T-47D. ( B ) Individual scattered dot plots and linear regression modelling of vMET1-Fc binding with respect to receptor density, p-MET, total MET protein, MET RNA, and MET CN. Pearson correlation coefficient (R) was calculated for each variable comparison. Strong correlation was observed between antibody binding and MET receptor density. ( C ) Viability assay to assess proliferation with concentrations of vMET1-Fc ranging from 1 fM to 1 μM does not show impact on cell survival across indicated cell lines. Points mean; bar SEM (n = 6). ( D ) Sensorgram showing the binding of HGF to sensors loaded with human MET protein, and then dissociating in assay buffer (top) and the same assay, but vMET1-Fc is allowed to associate with sensors before the addition of HGF (bottom).

Article Snippet: MET copy number variation was determined by predesigned TaqMan Copy Number Assay (Life Technologies, Hs04993403_cn; FAM-dye labeled) and TaqMan Copy Number Reference Assay as RNaseP (Life Technologies, #4403328, VIC-dye labeled). qPCR was performed using QuantStudio3 Real-Time PCR System (Applied Biosystems).

Techniques: Expressing, Binding Assay, Flow Cytometry, Comparison, Viability Assay

Journal: bioRxiv

Article Title: A MET-Targeted Variable New Antigen Receptor (VNAR) Theranostic for Non-Small Cell Lung Cancer

doi: 10.64898/2026.01.30.702875

Figure Lengend Snippet: ( A ) Aggregate data from high-content live-cell imaging of vMET1-Fc internalization into MET-expressing EBC-1 and UW-Lung-21 cells and MET-negative T-47D cells. Antibody was directly labelled with pH-sensitive pHrodo Red, which increases fluorescence with decreasing pH, and resulting fluorescence was measured as integrated signal intensity normalized to confluency for three days. Points mean; bar SEM (n =5). ( B ) Representative confocal microscopy images assessing vMET1-Fc internalization into MET-positive and -negative cell lines over time. Nuclei (blue), cell membranes (red), endosomes (green) and vMET1-Fc (white) are stained in large composite images, while separated channels are shown in smaller images. Blue line (inset) shows axis of the graph depicting vMET1-Fc and endosome signal at each time point. ( C ) Quantification of percent overlap of vMET1-Fc color channel with endosomal color channel per cell (approximately 100 cells per time point per cell line) affirms internalization observed only within MET-expressing cell lines.

Article Snippet: MET copy number variation was determined by predesigned TaqMan Copy Number Assay (Life Technologies, Hs04993403_cn; FAM-dye labeled) and TaqMan Copy Number Reference Assay as RNaseP (Life Technologies, #4403328, VIC-dye labeled). qPCR was performed using QuantStudio3 Real-Time PCR System (Applied Biosystems).

Techniques: Live Cell Imaging, Expressing, Fluorescence, Confocal Microscopy, Staining

Journal: bioRxiv

Article Title: A MET-Targeted Variable New Antigen Receptor (VNAR) Theranostic for Non-Small Cell Lung Cancer

doi: 10.64898/2026.01.30.702875

Figure Lengend Snippet: ( A ) Representative images from PET/CT scans of mice bearing xenografted tumors of EBC-1, UW-Lung-21, or T-47D cells (n = 4 per cell line) injected with [ 89 Zr]Zr-vMET1-Fc and imaged at the given intervals. ( B ) Region of interest (ROI) analysis of the PET images quantified radioactivity uptake (injected dose per gram (%ID/g)) across all time points for the heart and tumor. ( C ) Ex vivo biodistribution analysis of [ 89 Zr]Zr-vMET1-Fc activity across organs and tissue collected at 96 h post injection. There is significantly higher measured activity in MET-altered tumors compared to MET-negative tumors. Values are the mean (n = 4) ± SEM.

Article Snippet: MET copy number variation was determined by predesigned TaqMan Copy Number Assay (Life Technologies, Hs04993403_cn; FAM-dye labeled) and TaqMan Copy Number Reference Assay as RNaseP (Life Technologies, #4403328, VIC-dye labeled). qPCR was performed using QuantStudio3 Real-Time PCR System (Applied Biosystems).

Techniques: Positron Emission Tomography-Computed Tomography, Injection, Radioactivity, Ex Vivo, Activity Assay

Journal: Materials Today Bio

Article Title: Co-delivery of panobinostat and siSTAT3 using engineered M1 exosomes to establish a one-two punch therapeutic strategy for glioblastoma recurrence

doi: 10.1016/j.mtbio.2025.102680

Figure Lengend Snippet: Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, and FBS free medium with or without fMLP (100 μM, MCE, HY-P0224) was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.

Article Snippet: Monolayered HUVEC with TEER values greater than 300 Ω cm 2 were adopted as the BBB model. Then, 0.2 μg/μL of PKH67-labeled iM1-EXOs-PAN was added to the upper chamber, and FBS-free medium with or without fMLP (100 μΜ, HY-P0224, MCE) was added to the lower chamber [ ].

Techniques: In Vitro, Confocal Microscopy, Co-Culture Assay

Journal: bioRxiv

Article Title: Cancer-Associated Mesothelial Cells Drive Immune Escape and Therapy Resistance in Ovarian Cancer

doi: 10.64898/2026.01.07.698232

Figure Lengend Snippet: A) Quantitative PCR (qPCR) analysis of Upk3b and Serpinb2 expression in immortalized mesothelial cells (iMC) stimulated with KPCA-conditioned media and Met5a stimulated with patient D HGSOC media (ptD) or Ov90 ovarian cancer cell line in a time-dependent manner (0-96hrs). B) Western blot analysis of UPK3B and SERPINB2 expression in iMC treated with KPCA-condioned culture medium at 0h, 24h, 48h, 72h and 96h (left panel) and Met5A stimulated with conditioned media from ptD/Ov90 cell-lines versus unstimulated controls at 72h (right panel). C) Immunofluorescence analysis of UPK3B and SERPINB2 in iMC and Met5A comparing conditions stimulated with either KPCA-conditioned media or ptD/Ov90 media versus unstimulated controls at 72 h. D) Immunofluorescence staining for ACTA2 and COL4A1 in both primary MCs and immortalized MCs (iMCs) after 96 hours of exposure to KPCA-conditioned media

Article Snippet: The Met5a cell line was purchased from ATCC and cultured in DMEM medium supplemented with 5% FBS.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunofluorescence, Staining

Journal: Proteomes

Article Title: Proteomics and Bioinformatics Profiles of Human Mesothelial Cell Line MeT-5A

doi: 10.3390/proteomes14010002

Figure Lengend Snippet: Summary of cellular and conditioned medium proteomes of MeT-5A. ( a ) Venn diagram illustrating the total number of proteins identified by high-throughput LC-MS/MS from two distinct sample types: cellular lysate and conditioned cell culture medium. The numbers within each circle represent the total number of proteins uniquely identified in either the cellular lysate (blue circle) or the conditioned medium (green circle). The overlapping region indicates the number of proteins that were identified in both the cellular and conditioned medium samples. ( b ) Distribution of identified proteins (blue bars) relative to known total protein-coding genes (black bars) across human chromosomes. This bar plot illustrates the number of proteins identified in this study compared to the total number of protein-coding genes known for each individual chromosome. The x-axis represents the individual chromosomes, while the y-axis indicates the count of protein-coding genes or proteins. Blue bars denote the count of proteins identified in this study, while black bars represent the total number of protein-coding genes on each chromosome, according to Ensembl annotations.

Article Snippet: We cultured three technical replicates of MeT-5A cells (ATCC, Manassas, VA, USA) at 37 °C and 5% CO 2 in modified optimum medium.

Techniques: High Throughput Screening Assay, Liquid Chromatography with Mass Spectroscopy, Cell Culture

Journal: Proteomes

Article Title: Proteomics and Bioinformatics Profiles of Human Mesothelial Cell Line MeT-5A

doi: 10.3390/proteomes14010002

Figure Lengend Snippet: Dynamic range of cellular and conditioned medium proteomes of MeT-5A. ( a , b ) depict the wide dynamic range of proteins present in MeT5A cell lysate and cell conditioned medium, respectively, spanning seven orders of magnitude as shown by protein mean normalized abundance (PSM) on the Y-axis. The X-axis represents individual protein rank based on abundance. The abundance of each protein estimated based on the spectral count (defined by the total number of identified peptide spectra matched (PSM) to the protein of interest, including those redundantly identified) revealed the typical S-shaped distribution over the seven orders of dynamic range of MS signals. ( c , d ) show top ten most abundant proteins in the cell lysate and conditioned medium, respectively.

Article Snippet: We cultured three technical replicates of MeT-5A cells (ATCC, Manassas, VA, USA) at 37 °C and 5% CO 2 in modified optimum medium.

Techniques: