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 事实证明，遗传改性的小鼠模型对于抗体相关的产生至关重要 生物药物（生物制剂）。迄今为止，大多数生物制剂源自小鼠模型和小动物 模型不仅用于生成抗体，而且还用作进一步优化和测试的平台 生物制剂。基于骆驼的抗体，具有优质的抗原结合和物理化学 特性（稳定性，亲水性等）并未意识到它们的全部潜力，就像 常规抗体具有。这建立在骆驼的物流和财务障碍免疫中 单克隆重链抗体（HCAB）生产的姿势以及体外技术不能 完全概括出极度多样化的高亲和力粘合剂的特殊自然选择 发生在动物中。在这个SBIR项目中，我们建议在鼠宿主中开发遗传平台 发现和开发部分人性化的杂种HCAB（及其产品）。自从他们发现 1990年代初，HCAB对生物技术，诊断和治疗领域产生了渐进兴趣 由于它们的内在特性和适应性。除了小尺寸和稳健性和优越性 访问困难的表位，HCAB可以轻松处理并用作单个域绑定单元 （VHH）在保留对抗原的亲和力（与常规抗体相反）。提议 携带工程免疫球蛋白基因座的目标小鼠模型将增强高高的生产 亲和力HCAB通过作为替代性的混合AB宿主。它将允许自然的体内亲和力成熟 小动物平台中的抗原特异性HCAB，可用于进一步的遗传操作。它将启用 比天然骆驼宿主大的队列大小，并简化了HCAB的生成，从而进一步提供 为下游应用开发HCAB和VHH域的潜力。在我们的目标1中，我们专注于 通过添加更多的骆驼VHHS和 将改良的人VHS引入基因座，同时还评估B细胞开发和抗体Affininty 和多样性。在AIM 2中，我们的重点是用AB响应的曲目和效率对 竞争技术通过使用与疾病相关的，困难的抗原和进展有望击中杂交瘤 开发和大规模抗体产生。在实现这些基于里程碑的目标时，我们将 能够开发我们的业务并开始对单个实验室的平台许可并建立 制药公司支持发现针对高价值靶标的新型抗体。