Multifaceted mutational immunotherapeutic approach to design therapeutic mAbs to combat monkeypox disease via integrated screening algorithms and antibody engineering

Abstract

After a multi-country outbreak, the monkeypox (MPX) disease was designated a global public health emergency on July 23, 2022. Some antiviral medications tailored to the smallpox virus are currently being used to treat the disease. There is no specific treatment for the MPX disease with minimal negligible side effects. The engineering of antibodies has increased dramatically since the US Food and Drug Administration (US FDA) approved the first therapeutic monoclonal antibody (mAb) in 1986. mAbs have revolutionized biomedical research and have been used with remarkable precision for avoiding undesirable consequences. So, in this study, mAbs from the Thera-SAbDab (Therapeutic Structural Antibody Database) were screened using the ClusPro protein–protein docking server against the critical enzymes of the monkeypox virus (thymidine kinase, methyltransferase, D9 decapping enzyme, and RNA polymerase). Based on the predicted ClusPro docking score, binding affinity (ΔG), dissociation constant (K_d), and physiochemical properties, the best two mAbs (eculizumab and vofatamab) were designated for further investigation. Furthermore, the CUPSAT server and PyMol mutagenesis wizard were employed to generate a mutant pool (up to triple mutant through permutation combinations) and investigate the binding affinity of the candidate mAbs following point mutation. Eventually, the mAbs eculizumab mutant (L: L92F) and vofatamab mutant (L: H94T, L: Q96V) were identified as the most effective and promising inhibitors targeting all four MPXV enzymes, based on molecular dynamics (MD) simulations and MD trajectory assessment. In the future, in vitro and in vivo experiments on promising mAbs identified and developed by us could aid virus neutralization in MPXV-infected patients.