​In Silico Design of a Bispecific DuoBody Antibody Targeting S100A4 and T-Cell Receptors to Enhance Immune Infiltration in Glioblastoma

Glioblastoma (GBM) is the most aggressive and lethal primary brain tumor in adults, characterized by rapid progression and resistance to current therapies. The S100A4 protein, secreted by GBM cells, plays a critical role in tumor invasion, metastasis, and immune evasion. To address these challenges, this study focuses on the in silico design of a bispecific DuoBody antibody capable of simultaneously binding to the S100A4 protein and T-cell receptors (TCRs). By bridging T cells directly to the tumor microenvironment, the designed antibody aims to enhance immune infiltration and antitumor activity. Using computational tools such as AlphaFold 3 for structure prediction, P2Rank and ScanNet for binding site identification, HDOCK for molecular docking, and PLIP/PRODIGY for interaction and binding energy analysis, we evaluated the structural feasibility and binding affinity of candidate antibody complexes. Preliminary results suggest that antibody 1IGT demonstrated the strongest interaction with S100A4, supporting its potential as a therapeutic candidate. This work highlights the promise of DuoBody antibodies as next- generation immunotherapeutics for GBM, though experimental validation remains necessary.