An integrated multi-scale camouflaging platform for cloaking immunogenicity and evading non-specific clearance of therapeutic proteins

Monoclonal antibodies (mAbs) offer a significant opportunity in the prophylaxis and therapy of diseases caused by biological and chemical warfare agents. Many of the mAbs developed against these threats have non-human origins (e.g., mouse, rat), which are associated with undesirable properties including short serum persistence and elicitation of human anti-drug antibodies (ADAs). Protein engineering is commonly used to reduce immunogenicity (i.e., by introducing more humanness into non-human sequences) and improve pharmacokinetics (i.e., by enhancing neonatal Fc receptor (FcRn) binding). However, the sequence changes required to achieve these objectives are often deleterious to antigen binding and/or Fc effector functions. Moreover, many human/humanized mAbs still trigger ADA responses and infusion reactions. To address these issues, the proposed work seeks to develop a multi-scale camouflaging platform for optimizing the pharmacokinetic (PK) behavior of therapeutic proteins. Specifically, a panel of non-human mAbs will be camouflaged using a suite of novel protein design/engineering/glycosylation strategies for masking the major modes of immunological molecular recognition and blocking the primary routes of non-specific clearance. Importantly, studies proposed here will integrate these objectives so as to engineer all simultaneously. We will work with PI Dr. Matthew DeLisa from Cornell University and our collaborators to achieve the overall project goals. Specifically, we will focus our efforts on the following tasks: (i) Develop methods to express antibodies in cell-free protein synthesis; and (ii) Develop customized mAb glycosylation, whereby methods for designer glycan construction and site-specific protein glycosylation are used.