Antibody Optimization Core

抗体优化核心

• 批准号: 10402341
• 负责人: Galit Alter
• 金额: $ 69.29万
• 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 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10402341
• 关键词: 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; Data Analyses; Effectiveness; Effector Cell; Engineering; Ensure; Enzymes; Epitopes; Evolution; Family; Filovirus; Future; Generations; Genomics; Geometry; Goals; Health; Human; Immune; Immunity; Immunoglobulin Constant Region; Immunotherapeutic agent; Infection; Innate Immune System; Institutes; Knock-in; Knock-out; Lassa virus; Libraries; Link; Measures; Mechanics; Mediating; Methods; Mission; Modification; Monoclonal Antibodies; Mutation; Performance; Point Mutation; Polysaccharides; Research Project Grants; Serology; Site; Standardization; Surface; System; Technology; Therapeutic; Therapeutic Monoclonal Antibodies; Time; Treatment Efficacy; Universities; Vaccines; Variant; Viral; Virus; Virus Diseases; Washington; Work; Yeasts; antibody engineering; antigen binding; base; biodefense; data sharing; efficacy testing; emerging pathogen; falls; glycosylation; immune function; improved; interest; lentiviral-mediated; natural antibodies; neutralizing antibody; 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.

核B（抗体优化核心） 抽象的 在没有保护性疫苗和已知免疫相关的情况下，单克隆的被动转移 抗体（mAb）是针对，控制和清除感染的有前途的策略。虽然基因组测序 B细胞克隆技术彻底改变了我们几乎任何人都产生mab的图书馆的能力 感兴趣的目标，在许多情况下，这些mAb不足，并且无法实现足够的保护性免疫力 临床或临床研究。幸运的是，新的抗体工程策略现在使我们能够增强mab- 通过优化MAB功能介导的保护性免疫。 mAb抗原结合的演变 （FV）和恒定区域（FC）可以改善整体MAB性能。该核心，抗体优化核心 （核心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 功能以及工程方法以优化FAB和FC MAB域的有效性。 该优化将通过：（i）使用酵母表面显示来发展抗原的FV亲和力 目标； （ii）已知会改变对FCG受体（FCGR）和中的亲和力的FC结构域中的点突变的产生 转弯会影响抗体依赖性细胞介导的细胞毒性）； （iii）通过表达在FC糖基化中改变 具有（a）CRISPR/CAS9介导的基因敲除或（b）慢病毒介导的酶敲除酶的专门细胞系 消除碳水化合物的碳水化合物合成或（c）废除核心构成的（c）点突变。我们将 评估使用两个高通量系统的工程mAB介导的一系列免疫功能 血清学和基于细胞的测定法，并与所有三个研究项目合作以确定 在细胞培养和动物模型中优化针对真实病毒的抗体。核心B还将集成 使用核心C（计算分析和建模核心）来解释我们的结果并揭示 由项目确定的MAB特征和保护措施。我们将与核心A合作（管理 核心）以确保所有项目和其他利益相关者之间的数据策划和共享。在一起，我们会的 发现并提供优化的，高效的MAB疗法，以针对三个主要的病毒系列。我们 进一步寻求了解哪些发现是一般规则，哪些策略是特定于每个病毒家族的， 提供尚不存在的抗体选择和优化的系统，并建立广泛的适用 也针对新型病原体的MAB发现和分娩平台。