Interdisciplinary protein engineering approach to design high affinity antibodies for flaviviruses

摘要 重组单抗被认为是抗病毒治疗的新模式 感染。开发病毒mAb的一个特殊挑战是分离和鉴定具有 针对单个病毒家族的多个成员的广谱中和活性(BNAbs)提供 免疫治疗优势，特别是对感染病原体来说，其病例可能迅速进展为 严重的疾病。 我们鉴定bNAbs的目标是通过协同结合 计算和实验方法，以增强特定单抗的广度和效力 精心设计的突变。噬菌体展示提供了一种高效的方式来随机浏览多达1010个独特的文库 成员同时加入。然而，典型的~15个残基的抗体-抗原界面呈现出 2015个突变变种的组合可能性，其中只有一小部分可以抽样。结果，噬菌体 展示和其他抗体组合方法依赖于氨基酸多样性或 界面位置，这限制了该方法的有效性。为了绕过这一限制，我们建议 开发一种跨学科的方法，其中以计算设计的、以残留物为基础的药效团 抗体-抗原界面的描述被用来指导后续噬菌体展示中的文库设计 实验。作为这种方法的试验台，我们将设计新型的抗黄病毒的bNAbs，包括 全球重要病原体登革热病毒和寨卡病毒。

Abstract Recombinant monoclonal antibodies considered a new modality of therapeutic treatment to fight viral infections. A particular challenge for viral mAb development is the isolation and characterization of mAbs with broadly neutralizing activity (bNAbs) against multiple members of a single viral family to provide an immunotherapeutic advantage, particularly for infectious agents whose cases can progress rapidly toward serious disease. Our goal for the identification of bNAbs is protein engineering by a synergistic combination of computational and experimental methods, to enhance the breadth and potency of a specific mAb using designed mutations. Phage display provides an efficient way to randomly explore up to 1010 unique library members simultaneously. However, a typical antibody-antigen interface of ~15 residues present a combinatorial possibility of 2015 mutational variants, only a fraction of which can be sampled. As a result, phage display and other antibody combinatorial methods are reliant on restriction of either amino acid diversity or interfacial positions, which limits the effectiveness of the approach. To circumvent this limitation, we propose to develop an interdisciplinary approach where computationally designed, residue-based pharmacophore descriptions of the antibody-antigen interface are used to direct the library design in subsequent phage display experiments. As a test bed for this method, we will engineer novel bNAbs against flaviviruses, which include the globally important pathogens dengue virus and Zika virus.