Computational approaches to design Anti-CD۵۲ Nano body: A new insight for B-CLL and MS therapeutic strategies

Backgrounds: The research in Nano bodies as therapeutics for cancer and autoimmune diseasehas gained attention because of their appreciable advantages compared to conventionalantibodies. Delivery of monoclonal antibodies to tumor cells in vivo is limited due to their largesize, while Nano bodies show excellent tissue penetration, are safe and of low immunogenicity inhumans, and can simply be engineered and produced in a prokaryotic host. Epitopes that areoften unreachable to conventional antibodies can be recognized by Nano bodies. Alemtuzumabis an approved mAb that is being indicated to treat B-cell chronic lymphocytic leukemia (BCLL)as well as the relapsing forms of multiple sclerosis (MS). In this study, we propose anapproach for designing and generating an anti-CD۵۲ Nano body.Materials and Methods: Complementarity-determining regions (CDRs) were grafted fromconventional antibody layouts onto Nano body frameworks to generate antigen binders andobtain the Nano body protein sequence, using the bioinformatics tools. The reverse translationwas done for obtained Nano body protein sequence, with the application of codon optimizing.We performed site-directed mutagenesis to enhance the antigen-antibody interaction affinity.The procedure was certified using in silico cloning of obtained Nano body gene sequence intothe prokaryotic host.Results: These findings propose that designing Nano bodies for targeted therapy of B-CLL andMS patients can have significant antitumor utility via enhanced affinity for the CD۵۲-bindingdomain and recognizing conserved epitopes. These epitopes are mainly inaccessible to humanmAbs.Conclusion: Although new targeted therapy approaches for cancer will continue to emerge,designing Nano bodies serve as a reassuring novel tool to help the patients.

کلیدواژه ها:

Complementarity-determining region (CDR) grafting ، B-CLL ، VHH ، Therapeutic antibodies ، Computational biology