Research and Development to Establish a Small Animal Model as a Significant Resource of High-Value Single-Domain Antibodies

建立小动物模型作为高价值单域抗体重要资源的研发

• 批准号: 10481556
• 负责人: Milen Kirilov
• 金额: $ 99.9万
• 依托单位: INGENIOUS TARGETING LABORATORY, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-03-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10481556
• 关键词: Affinity; Alpaca; Animal Model; Animals; Antibodies; Antibody Affinity; Antibody Formation; Antibody Response; Antigens; Autoantigens; B-Cell Development; Bacteria; Basic Science; Benchmarking; Binding; Biological; Biological Products; Biotechnology; Breeding; Businesses; Camels; Cells; Characteristics; Client; Clinical; Complex; Coupled; Custom; Development; Diagnostic; Diagnostics Research; Disease; Engineering; Enterobacter cloacae; Epitopes; Fab Immunoglobulins; Future; Gene Bank; Gene Proteins; Gene Targeting; Generations; Genes; Genetic; Genetic Engineering; Genetically Engineered Mouse; Goals; HIV; HIV Envelope Protein gp120; Homer 1; Housing; Human; Hybridomas; Hybrids; Immune; Immune response; Immunization; Immunize; Immunoglobulins; In Vitro; Individual; Laboratories; Licensing; Light; Light-Chain Immunoglobulins; Link; Llama; Logistics; Malignant Neoplasms; Medicine; Membrane; Modeling; Mus; Natural Selections; Nature; Outcome; Performance; Phage Display; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Play; Process; Production; Property; Proteins; Protocols documentation; Resources; Role; SARS-CoV-2 spike protein; Services; Shark; Small Business Innovation Research Grant; Targeted Research; Technology; Testing; Therapeutic; Therapeutic Agents; Virus; Yeasts; animal resource; antibody diagnostic; antibody libraries; antigen binding; base; clinical application; clinical diagnostics; cohort; cost; embryonic stem cell; environmental stressor; experimental study; flexibility; genetic manipulation; hybrid antibody; hydrophilicity; immunogenicity; improved; in vivo; in-vivo diagnostics; interest; mouse model; nanobodies; neutralizing antibody; novel; pathogen; pathogenic microbe; preservation; research and development; sensor; standard of care; validation studies; virus host interaction

Project Summary Genetically-modified mouse models have proven to be essential for the production of antibody-related biological drugs (biologics). To date, the majority of biologics originate from mouse models, and small animal models are used not only to generate the antibodies, but also as a platform for further optimization and testing of the biologics. Camelid-based antibodies, which have superior antigen binding and physicochemical properties (stability, hydrophilicity, etc.) have not realized their full potential, to the same extent that conventional antibodies have. This is founded in the logistic and financial hurdles immunization of camelids pose for monoclonal heavy-chain antibody (HCAb) production and the fact that in vitro technologies cannot fully recapitulate the exceptional natural selection towards extremely diversified, high-affinity binders that occurs in animals. In this SBIR project we propose to develop genetic platforms in a murine host for the discovery and development of partially humanized hybrid HCAbs (and their products). Since their discovery in the early 1990s, HCAbs have generated progressive interest in the biotech, diagnostic and therapeutic fields due to their intrinsic properties and adaptability. Apart from a small size paired with robustness and superior access to difficult epitopes, HCAbs can be easily processed into, and utilized as, single domain binding units (VHH) while preserving their affinity towards antigens (in contrast to conventional antibodies). The proposed targeted mouse models carrying an engineered immunoglobulin locus will potentiate the production of high affinity HCAbs by serving as an alternative, hybrid Ab host. It will allow natural, in vivo affinity-maturation of antigen-specific HCAbs in a small animal platform, one amenable to further genetic manipulation. It will enable larger cohort sizes than the natural camelid hosts, and streamline HCAb generation, thus providing further potential for the development of HCAb and VHH domains for downstream applications. In our Aim 1, we focus on honing and characterizing our hybrid camelid immunoglobulin locus by adding more camelid VHHs and introducing modified human VHs into the locus while also evaluating B-cell development and antibody affininty and diversity. In Aim 2, our focus is to benchmark the the repertoire and efficiency of the Ab response with competing technologies by using disease-relevant, difficult antigens and progress promising hits to hybridoma development and larger scale antibody production. In accomplishing these milestone based Aims, we will be able to develop our business and begin licensing of the platforms to individual labs and established pharmaceutical companies to support discovery of novel antibodies for high-value targets.

项目摘要 转基因小鼠模型已被证明是生产抗体相关抗体的关键。 生物药品（biologics）。迄今为止，大多数生物制剂来源于小鼠模型和小动物模型。 模型不仅用于生成抗体，还可作为进一步优化和测试的平台 的生物学。基于骆驼科动物的抗体，其具有上级抗原结合和物理化学性质， 性能（稳定性、亲水性等）没有充分发挥其潜力， 传统的抗体。这是建立在骆驼免疫的后勤和财政障碍 单克隆重链抗体（HCAb）的生产和体外技术不能 完全概括了异常的自然选择对极其多样化，高亲和力的粘合剂， 发生在动物身上。在这个SBIR项目中，我们建议在小鼠宿主中开发遗传平台， 部分人源化杂交HCAb（及其产物）的发现和开发。自从他们在1999年被发现以来， 在20世纪90年代早期，HCAb在生物技术、诊断和治疗领域引起了越来越多的兴趣 由于它们的固有特性和适应性。除了小尺寸搭配坚固耐用和上级 由于HCAb能够接近困难的表位，因此HCAb可以容易地加工成单结构域结合单元并用作单结构域结合单元 (VHH)同时保持它们对抗原的亲和力（与常规抗体相反）。拟议 携带工程化免疫球蛋白基因座的靶向小鼠模型将增强高免疫球蛋白的产生。 通过充当替代的杂合Ab宿主来亲和HCAbs。它将允许天然的，体内亲和力成熟， 抗原特异性HCAbs在小动物平台，一个适合进一步的遗传操作。它将使 比天然骆驼宿主更大的群体规模，并简化HCAb的产生，从而提供进一步的 HCAb和VHH结构域用于下游应用的开发潜力。在目标1中，我们专注于 通过添加更多的骆驼VHH来磨练和表征我们的杂交骆驼免疫球蛋白基因座， 将修饰的人VH引入基因座，同时还评估B细胞发育和抗体亲和力 和多样性。在目标2中，我们的重点是对抗体反应的库和效率进行基准测试， 通过使用疾病相关的、困难的抗原和进展有希望的杂交瘤命中来竞争技术 开发和大规模抗体生产。为了实现这些里程碑式的目标，我们将 能够发展我们的业务，并开始将平台许可给各个实验室， 制药公司，以支持高价值目标的新型抗体的发现。