Robust Antibody–Antigen Complexes Prediction Generated by Combining Sequence Analyses, Mutagenesis, In Vitro Evolution, X-ray Crystallography and In Silico Docking

摘要: Abstract Hu 15C1 is a potent anti-human Toll-like receptor 4 (TLR4) neutralizing antibody. To better understand the molecular basis of its biological activity, we used multidisciplinary approach to generate an accurate model 15C1–TLR4 complex. By combining site-directed mutagenesis, in vitro antibody evolution, affinity measurements and X-ray crystallography Fab fragments, identified key interactions across interface. These contact points were as restraints predict structure region bound TLR4 using computational docking. This was further evaluated validated by additional mutagenesis studies. The predicted complex indicates that antagonizes dimerization necessary for activation. study exemplifies how iterative cycles engineering can facilitate discovery components antibody-target interactions.