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

Loyau J, et al., 427(16):2647-62, J Mol Biol , 2015

This is a detailed investigation on structural and mechanistic elements contributing to the interaction between Hu15C1 neutralizing antibody and Toll-like receptor 4 (TLR4). The authors used a combined approach consist of site-directed and combinatorial mutagenesis along with X-ray crystallography and affinity measurements to dissect the molecular basis of this interaction. The information obtained from these experiments along with computational molecular docking lead to a mechanistic model. The model predicts that the antibody binding interferes with dimerization of the receptor, which is a crucial step for its activation. Both SPR and BLI systems were used for the kinetic characterization of the interaction. Epitope binning experiments were carried out using a Pall ForteBio Octet RED96 system equipped with Protein A biosensor tips.

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