Development and Optimization of Therapeutic Analogues of Anti-TNFα Antibody Infliximab

Previously, we have reported the crystal structures of Fab fragment of Infliximab in complex with TNF alpha. The structurally identified epitope on TNF alpha revealed the mechanism of TNF alpha inhibition by partially overlapping with the TNF alpha-receptor interface and the possibility to optimize the binding affinity. In this study, we launched a screen of a phage display library to isolate novel anti-TNF alpha antibodies based on the infliximab epitope. To develop novel anti-TNF alpha antibodies, structural analysis, the phage display antibody isolation, step by step antibody optimization, CDR residues random mutagenesis, and binding affinity characterization were performed. One of the novel antibodies generated on the backbone of infliximab, Inf3D6, has the superior binding affinity to TNF alpha, thus, demonstrating the potential for structure guided optimization for improvement of existing antibody-based therapeutics.