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