ArticleAntibody therapeutics2026
A novel bispecific antibody targeting OX40L and TNFα for the targeted treatment of rheumatoid arthritis.
Article in Antibody therapeutics, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Background: Rheumatoid arthritis is driven by T cell dysregulation and TNFα-mediated inflammation. Dual targeting of OX40 ligand (OX40L) and tumor necrosis factor alpha (TNFα) may offer synergistic therapeutic effects. Methods: A murine monoclonal antibody against OX40L (KD-0025) was generated and humanized via complementarity-determining region grafting. A bispecific construct incorporating single-chain variable fragments for OX40L and TNFα was expressed in CHO-K1 cells. High-expressing clones were selected, and antibodies were purified using Protein A/G chromatography. Binding to 293 T-OX40L cells, A172 cells, and TNFα was assessed by flow cytometry and ELISA. Functional blockade of OX40L-OX40 and TNFα-TNFR pathways was evaluated using a luciferase reporter assay. Binding kinetics were measured using the ForteBio Octet system. Results: The parental KD-0025 antibody showed EC50 values of 0.4024 μg/ml (human OX40L) and 0.1126 μg/ml (monkey OX40L), with an IC50 of 0.07893 μg/ml. Humanized KA-3966 improved EC50 (0.1374 μg/ml) and had an IC50 of 0.5997 μg/ml. The bispecific antibody KA-5484 demonstrated superior affinity (KD = 1.90 × 10 Conclusion: A humanized bispecific antibody targeting OX40L and TNFα was successfully developed, exhibiting potent dual-target engagement and blockade. The antibody demonstrated synergistic modulation of immune and inflammatory pathways, providing a promising foundation for future preclinical development. Highlights: The novel bispecific KA-5484 binds both OX40L and TNFα with high affinity.Dual targeting of OX40L and TNFα enables synergistic immune modulation.KA-5484 outperforms control antibodies in binding and inflammatory assays.KA-5484 shows superior efficacy in mitigating arthritis in the collagen antibody-induced arthritis mouse model.Enhanced cross-species reactivity supports the translational relevance of KA-5484.
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