humanized monoclonal igg1 antibody  (Genentech inc)

Journal: bioRxiv

Article Title: Molecular basis for the increased membrane fusion activity of the Ebola virus glycoprotein A82V variant from the 2013-2016 epidemic: insights from simulations and experiments

doi: 10.1101/2024.10.28.620686

Figure Lengend Snippet: ( A ) The GP ectodomain structure with residues in one protomer colored according to the difference in RMSF between the A82 and V82 simulations (ΔRMSF = RMSF A82 – RMSF V82 ). Key structural elements are indicated: RBS, receptor-binding site; α1 helix; HR1, heptad repeat helix 1. ( B ) ΔRMSF for each residue in the GP ectodomain. The data are presented as the average of each protomer across the triplicate simulations, with error bars reflecting the standard error. ( C ) Visualization of the α1 helix in GP1 from representative frames in the A82 (cyan) and V82 (grey) simulations, indicating the loss of helical content in the V82 simulation. The region shown corresponds to the red dashed rectangle in panel (A). Quantification of the helical content of the α1 helix over the duration of the ( D ) A82 and ( E ) V82 simulations. Data are presented as the average of each protomer across the triplicate simulations, with error bars reflecting the standard error. ( F ) Box plots indicating the median, quantiles, and range of helical content seen across the simulations (A82, cyan; V82, grey). The p -value was determined by one-way ANOVA.

Article Snippet: The humanized anti-GP1 H3C8 IgG monoclonal antibody was expressed in ExpiCHO-S cells using ExpiFectamine CHO Reagent (ThermoFisher) according to the manufacturer’s standard expression protocol and a 2 to1 ratio of H3C8 Igκ to Igγ1 expression vectors.

Techniques: Binding Assay, Residue

Journal: NPJ Precision Oncology

Article Title: CCL3 predicts exceptional response to TGFβ inhibition in basal-like pancreatic cancer enriched in LIF-producing macrophages

doi: 10.1038/s41698-024-00742-3

Figure Lengend Snippet: A qPCR of Tgfb1 in RAW264.7 macrophages in presence or absence of rCcl3, after treatment with DMSO or 5 μM maraviroc. Results show mean ± s.d ( n = 3). B qPCR of Tgfb1 in RAW as single cultures or co-cultured with RC416, DT4313 or PAN610 transduced as indicated, after treatment with DMSO or maraviroc (5 μM). Results show mean ± s.d ( n = 3). C Representative western blot of pSmad2 and Smad2 in RAW264.7 in presence or absence of rCcl3, and as single cultures or in co-culture with RC416, DT4313 and PAN610 transduced as indicated, after treatment with DMSO or 5 μM maraviroc. Hsp90 was used as loading control. D Schematic representation of Tgfb1 promoter (available at UCSC Genome Browser GRCm38/mm10) with the position of ChIP probes (gray double arrowhead) and consensus Stat3 binding sites (TTC(N)2-4GAA) (vertical slashes) relative to the transcription starting site (TSS). E ChIP-qPCR of Stat3 occupancy at the Tgfb1 promoter in RAW264.7 macrophages upon stimulus with rCcl3, in the presence or absence of 5 μM maraviroc. The y -axis represents relative promoter enrichment, normalized on input material. IgG was set to 1. Data are represented as mean ± s.d. ( n = 4). F Representative western blot of secreted Ccl3 in the conditioned media of RAW264.7 macrophages stimulated or not with rCcl3, and treated with 5 μM maraviroc, 5 μM galunisertib, their combination or DMSO as control. G qPCR of Tgfb1 in RAW264.7 macrophages in the presence or absence of rCcl3 and treated as indicated. Results show mean ± s.d ( n = 3). H Secreted levels of Tgfβ1 ligand measured in conditioned medium of RAW264.7 macrophages treated as indicated using Enzyme-linked immunosorbent assay (ELISA). Data are expressed as mean ± s.d. ( n = 3). I Representative western blot of pSmad2 and Smad2 in RAW264.7 macrophages treated as indicated. Hsp90 was used as loading control. J , K qPCR of M1 ( J ) and M2 ( K ) markers in RAW264.7 macrophages in presence or absence of rCcl3 and treated as indicated. Results show mean ± s.d ( n = 3). P values in ( A , B , E , G , H , J and K ) were calculated by one-way ANOVA and Tukey’s test. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: In August 2023, Novartis discontinued the development of NIS793, a human IgG2 monoclonal antibody that binds to TGFβ1 and 2 ligands , which was under investigation in a randomized, placebo-controlled phase 3 trial in combination with gemcitabine and nab-paclitaxel for the first-line treatment of patients with metastatic PDAC (NCT04935359).

Techniques: Cell Culture, Western Blot, Co-Culture Assay, Control, Binding Assay, Enzyme-linked Immunosorbent Assay