Anti-IL-10 Antibody Humanization by SDR Grafting with Enhanced Affinity to Neutralize the Adverse Response of Interleukin-10

The overexpression of IL-10 is associated with numerous diseases like lymphoma, melanoma, systemic lupus erythematosus, systemic sclerosis, and bullous diseases. To date, no therapeutic humanized monoclonal antibody has been reported to neutralize the harmful effect of overexpressed IL-10. JES3-9D7 is a diagnostic mouse monoclonal antibody that cannot be utilized to treat humans due to the generation of HAMA (Human anti-mouse antibody) and allergenicity. To counteract the hyperactive response of IL-10, here we utilized mutational immunotherapeutic approaches to humanize JES3-9D7 by the SDR (specificity-determining residues) grafting. The SDR residues of 9D7 were evaluated and grafted on the IgG1 using the PyMOL mutagenesis tool. The binding affinity, humanness score, toxicity, and allergenicity of this humanized anti-IL-10 antibody were estimated. A mutational approach was used to improve the binding affinity of this humanized anti-IL-10 antibody, and the stability and free energy of various mutations were assessed. Seven mutants were chosen based on Z-score to prepare multiple mutant libraries. Molecular Dynamics Simulations were used to examine the stability and thermodynamic properties of two lead mutants. As a result, these humanized mutant antibodies could be investigated as potential lead therapeutics against IL-10 overexpression.