Interdisciplinary protein engineering approach to design high affinity antibodies for flaviviruses

跨学科蛋白质工程方法设计黄病毒高亲和力抗体

基本信息

批准号：
10294224
负责人：
Andras Fiser
金额：
$ 41.75万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-11-15 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10294224
关键词：
Address Algorithms Alphavirus Amino Acids Animals Antibiotics Antibodies Antibody Affinity Antibody Therapy Antigen-Antibody Complex Antigens Autoimmune Diseases B cell repertoire Beds Communicable Diseases Cytomegalovirus Dengue Virus Development Disease Ebola Effectiveness Engineering Evaluation FDA approved Family Filovirus Flavivirus Generations Goals HIV HIV-1 Homology Modeling Human Immunization Immunotherapeutic agent Infection Infectious Agent Influenza Libraries Lymphocyte Malignant Neoplasms Medical Methods Modality Monoclonal Antibodies Mutation Phage Display Phase I Clinical Trials Phase II Clinical Trials Positioning Attribute Protein Engineering Proteins Rabies Randomized Recombinants Sampling Serotyping Structural Models Structure Technology Testing Therapeutic Variant Viral Viral Hemorrhagic Fevers Virus Virus Diseases ZIKA Zika Virus antibody engineering base combinatorial cross reactivity design experimental study fighting human disease interdisciplinary approach interfacial member molecular dynamics mutant neutralizing antibody novel pathogen pharmacophore preference response screening

项目摘要

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.

抽象的重组单克隆抗体被认为是对抗病毒的新治疗方式感染。病毒单克隆抗体开发的一个特殊挑战是单克隆抗体的分离和表征针对单个病毒家族的多个成员的广泛中和活性（bNAb），以提供免疫治疗优势，特别是对于其病例可以迅速进展的传染源严重的疾病。我们鉴定 bNAb 的目标是通过协同组合进行蛋白质工程计算和实验方法，使用以下方法增强特定 mAb 的广度和效力设计的突变。噬菌体展示提供了一种随机探索多达 1010 个独特库的有效方法成员同时。然而，典型的约 15 个残基的抗体-抗原界面呈现出 2015 年突变变体的组合可能性，只能采样其中的一小部分。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，限制了该方法的有效性。为了规避这个限制，我们建议开发一种跨学科方法，通过计算设计、基于残留的药效基团抗体-抗原界面的描述用于指导后续噬菌体展示中的文库设计实验。作为该方法的测试平台，我们将设计针对黄病毒的新型 bNAb，其中包括全球重要的病原体登革热病毒和寨卡病毒。