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Journal: bioRxiv
Article Title: De novo masking domains that gate TNF-family ligand assembly and activity
doi: 10.64898/2026.04.20.719557
Figure Lengend Snippet: The antibody formats shown in were conjugated with MMAE, and cell binding and cytotoxicity were evaluated in HER2-positive SK-BR-3 cells. Protein concentrations are reported per molecular species (monomeric or trimeric ADC, as indicated). Where indicated, trimeric species were re-isolated by SEC before downstream assays. a, Proposed mechanism for the switchable ADC concept. Before activation, the construct is predominantly monomeric with reduced apparent binding and low payload density. Protease-triggered activation promotes assembly into a trimeric state with increased functional valency and complex-level payload density, which may enhance target-cell killing. b, Binding of monomeric and trimeric ADC species to HER2-positive SK-BR-3 cells measured by cell-based ELISA. Data are shown as mean ± s.d. from n = 3 technical replicates. c, Cytotoxic activity of ADCs in HER2-positive SK-BR-3 cells. Cell viability (%) was calculated as 100 × S/V, where S is the sample signal and V is the 100% viability control (cells plus medium). Data are shown as mean ± s.d. from n = 3 technical replicates.
Article Snippet: Cytotoxic activity of
Techniques: Binding Assay, Isolation, Activation Assay, Construct, Functional Assay, In-Cell ELISA, Activity Assay, Control