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: The Journal of Biological Chemistry

Article Title: An RNAi-based high-throughput screening assay to identify small molecule inhibitors of hepatitis B virus replication

doi: 10.1074/jbc.M117.775155

Figure Lengend Snippet: A reversion assay in HepG2 cells was used as a drug screening platform. A, immunofluorescence analysis and schematic representation of plasmid constructs used for the generation of the GFP-expressing HepG2 line, GFP-shRNA RNAi sensor line, GFP-reverted HepG2 cells that overexpressed HBx, and re-silenced GFP reverted cell treated by small molecule (IR415). B, real-time RT-PCR analysis of GFP transcript to show relative abundance of GFP in different cell lines: HepG2 cells with no GFP, HepG2/GFP cells (dark green) and HepG2/GFP-shRNA RNAi sensor line (light green). C, dot plot analysis from FACS data shows GFP expression in different HepG2 cell lines. Plots depict forward versus side scatters (SSC) for the expression of the GFP reporter at wavelength (488) with FL1 detector (530/40). D, histogram showing levels of GFP expression in HepG2 cell lines transfected with HBx following exposure with IR415 along with suitable controls. E, graphical representation of concentration-dependent denomination of liner suppression activity of IR415 at 50 μm, 100 μm and 200 μm in HepG2/GFP-shRNA line transfected with HBx. The error bars are the standard deviation of the mean of the ordinate values.

Article Snippet: Generation of HepG2 RNAi sensor line HepG2 cells were transfected with 6 μg of plasmid construct using pGFP-V-RS series of vectors expressing tGFP (OriGene, Rockville, MD).

Techniques: Immunofluorescence, Plasmid Preparation, Construct, Expressing, shRNA, Quantitative RT-PCR, Transfection, Concentration Assay, Activity Assay, Standard Deviation

Journal: The Journal of Biological Chemistry

Article Title: An RNAi-based high-throughput screening assay to identify small molecule inhibitors of hepatitis B virus replication

doi: 10.1074/jbc.M117.775155

Figure Lengend Snippet: HBV antigen kinetics and pre-genomic RNA level upon drug exposure are shown. A and B, the level of HBeAg and HBsAg was measured at different time points. Antigen titers were counted for HepG2 cells transfected with full-length pHBV plasmid that contained 3.5 kb HBV genome. Tenofovir is considered as positive control and DMSO as negative control. 24/24 denotes 24-h incubation of pHBV and 24-h treatment with IR415. 48/48 h indicates 48-h incubation of pHBV for viral proliferation and 48-h exposure post drug treatment. C, concentration-dependent inhibition constrains of IR415 have been significant for both HBeAg and HBsAg production during both 24- and 48-h exposure. D, the level of pre-genomic RNA of HBV post drug exposure. Log2 value of fold change for pre-genomic RNA is in reference to HBV transfected control cell line. The mean levels (± S.D.) were calculated from three replicate transfections. IU/ml measures the antigen titer for assessment of the analytical sensitivity of HBeAg assay which indicates active viral replication and infectivity.

Article Snippet: Generation of HepG2 RNAi sensor line HepG2 cells were transfected with 6 μg of plasmid construct using pGFP-V-RS series of vectors expressing tGFP (OriGene, Rockville, MD).

Techniques: Transfection, Plasmid Preparation, Positive Control, Negative Control, Incubation, Concentration Assay, Inhibition, Infection