Antibody Optimization Core

抗体优化核心

• 批准号: 10617741
• 负责人: Galit Alter
• 金额: $ 62.43万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10617741
• 关键词: Acceleration; Address; Affect; Affinity; Alphavirus; Animal Model; Antibodies; Antibody Affinity; Antigen Targeting; Antigens; B-Lymphocytes; Binding; Biological Assay; CRISPR/Cas technology; Carbohydrates; Cell Culture Techniques; Cell Line; Cell physiology; Cells; Clinical Research; Clone Cells; Communicable Diseases; Computer Analysis; Computer Models; Data; Effectiveness; Effector Cell; Engineering; Ensure; Enzymes; Epitopes; Evolution; Family; Fc domain; Filovirus; Future; Generations; Genomics; Geometry; Goals; Health; Human; Immune; Immunity; Immunoglobulin Constant Region; Immunotherapeutic agent; Infection; Innate Immune System; Knock-in; Knock-out; Lassa virus; Libraries; Link; Measures; Mechanics; Mediating; Methods; Mission; Modification; Monoclonal Antibodies; Mutation; Passive Transfer of Immunity; Performance; Point Mutation; Polysaccharides; Research Project Grants; Serology; Site; Standardization; Surface; System; Technology; Therapeutic; Therapeutic Monoclonal Antibodies; Time; Treatment Efficacy; Universities; Vaccines; Variant; Viral; Viral Physiology; Virus; Virus Diseases; Washington; Work; Yeasts; antibody and antigen binding; antibody engineering; biodefense; data sharing; efficacy testing; emerging pathogen; falls; glycosylation; immune function; improved; interest; lentiviral-mediated; natural antibodies; novel; pathogen; preclinical study; prevent; rational design; receptor; therapeutic candidate

CORE B (Antibody Optimization Core) Abstract In the absence of protective vaccines and known correlates of immunity, passive transfer of monoclonal antibodies (mAbs) is a promising strategy to target, control and clear infections. Although genomic sequencing and B cell cloning technologies revolutionized our capacity to generate libraries of mAbs against nearly any target of interest, in many cases these mAbs fall short, and fail to achieve sufficient protective immunity, in pre- clinical or clinical studies. Fortunately, new antibody engineering strategies now allow us to enhance mAb- mediated protective immunity through optimization of mAb functionality. Evolution of the mAb antigen-binding (Fv) and constant regions (Fc) can improve overall mAb performance. This Core, the Antibody Optimization Core (Core B) will support all three CETR Research Projects by providing high-throughput functional profiling of mAb functionality, as well as engineering approaches to optimize the effectiveness of both Fab and Fc mAb domains. This optimization will be engineered through: (i) use of yeast surface display to evolve Fv affinity for antigen targets; (ii) generation of point mutations in the Fc domain known to alter affinity for Fcg receptors (FcgR) and in turn affect antibody-dependent cell-mediated cytoxicity); and (iii) alterations in Fc glycosylation via expression in specialized cell lines with (a) CRISPR/Cas9-mediated knockout or (b) lentiviral-mediated knockin of enzymes governing carbohydrate synthesis, or (c) point mutations in the Fc domain that abolish core fucosylation. We will assess a range of immune functions mediated by the engineered mAbs using both high-throughput systems serology and cell-based assays, and work with all three Research Projects to determine performance of optimized antibodies against authentic viruses in cell culture and in animal models. Core B will also integrate with Core C (Computational Analysis and Modeling Core) to interpret our results and reveal correlations between mAb features and measures of protection determined by the Projects. We will work with Core A (Administrative Core) to ensure that data are curated and shared among all projects and other stakeholders. Together, we will discover and deliver optimized, highly efficacious mAb therapeutics against three major families of viruses. We further seek to understand what findings are general rules and what strategies are specific to each virus family, to provide systems for antibody choice and optimization that do not yet exist, and to build a broadly applicable platform for mAb discovery and delivery against novel pathogens as well.

CORE B（抗体优化核心） 摘要 在缺乏保护性疫苗和已知的免疫相关性的情况下，单克隆抗体的被动转移是不可能的。 单克隆抗体（mAb）是靶向、控制和清除感染的有希望的策略。虽然基因组测序 和B细胞克隆技术彻底改变了我们产生抗几乎任何 在许多情况下，这些mAb达不到目的靶点，并且不能实现足够的保护性免疫， 临床或临床研究。幸运的是，新的抗体工程策略现在允许我们增强mAb- 通过优化mAb功能介导保护性免疫。mAb抗原结合的演变 (Fv)而恒定区（Fc）可以提高mAb的整体性能。这个核心，抗体优化核心 (Core B）将通过提供mAb的高通量功能分析来支持所有三个CETR研究项目 功能，以及优化Fab和Fc mAb结构域的有效性的工程方法。 （i）使用酵母表面展示以进化Fv对抗原的亲和力 （ii）在已知改变对Fcg受体（FcgR）的亲和力的Fc结构域中产生点突变，以及在已知改变对Fcg受体（FcgR）的亲和力的Fc结构域中产生点突变。 反过来影响抗体依赖性细胞介导的细胞毒性）;和（iii）通过在 具有（a）CRISPR/Cas9介导的敲除或（B）慢病毒介导的酶敲入的特化细胞系 控制碳水化合物合成，或（c）Fc结构域中消除核心岩藻糖基化的点突变。我们将 使用两种高通量系统评估由工程化mAb介导的一系列免疫功能 血清学和基于细胞的检测，并与所有三个研究项目，以确定性能 在细胞培养物和动物模型中针对真实病毒的优化抗体。核心B还将整合 核心C（计算分析和建模核心）来解释我们的结果，并揭示 项目确定的mAb特性和保护措施。我们将与核心A（行政 核心），以确保数据在所有项目和其他利益相关者之间得到管理和共享。我们将共同 发现并提供针对三大病毒家族的优化、高效的mAb疗法。我们 进一步寻求了解哪些发现是一般规则，哪些策略是针对每个病毒家族的， 提供目前尚不存在的抗体选择和优化系统，并建立广泛适用的 该平台也是针对新型病原体的mAb发现和递送的平台。