Business Potential of a Novel Small Animal Model and its Single-Chain Antibodies

新型小动物模型及其单链抗体的商业潜力

• 批准号: 10087209
• 负责人: Milen Kirilov
• 金额: $ 5.5万
• 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 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10087209
• 关键词: Affinity; Alpaca; Animal Model; Animals; Antibodies; Antibody Diversity; Antibody Formation; Antigens; Bacteria; Binding; Biological; Biological Testing; Biotechnology; Businesses; Camels; Cells; Collaborations; Development; Diagnostic; Disease; Engineering; Epitopes; Generations; Genetic; Genetic Engineering; Genetically Engineered Mouse; Human; Hybridomas; Hybrids; Immune; Immune response; Immune system; Immunization; Immunoglobulins; Immunologics; In Vitro; Individual; Logistics; Malignant Neoplasms; Modeling; Modification; Mus; Natural Selections; Nature; Pharmaceutical Preparations; Phase; Play; Process; Production; Property; Proteins; Role; Small Business Innovation Research Grant; Technology; Testing; Therapeutic; Virus; antigen binding; base; cohort; design; embryonic stem cell; genetic manipulation; hydrophilicity; in vivo; innovation; interest; mouse model; novel; pathogen; preservation

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. Unmet Need: Camelid-based antibodies, which have superior antigen binding and physico- chemical properties (stability, hydrophilicity, etc.) have not realized their full potential, to the same extent that 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. Product: In this SBIR project we propose to develop a genetic platform in a murine host for the discovery and development of partially humanized hybrid HCAbs (and their products) containing camelid VHH domains – a combination not found in nature. 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. Significance: 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). Innovation: The proposed targeted mouse model carrying a pre-engineered alpaca/human 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 sequential genetic engineering of a targeted hybrid alpaca/human immunoglobulin locus in embryonic stem cells. In Aim 2, our focus is to characterize the generation of HCAbs originating from the immune system of the newly generated hybrid mouse model and to validate the applicability of our small animal platform. HCAbs derived from our novel mouse model will be subject to basic immunological analysis allowing us to characterize the repertoire and efficiency of the Ab response to model antigens (Phase I). If successful, we will utilize the newly generated animal model to produce HCAbs against disease-relevant, difficult antigens and progress to hybridoma development in further collaborations (Phase II). Beyond this, our platform, based on its design, will also allow further genetic modifications to take place involving antibody domains from other species and/or further changes to enhance HCAbs diversity, as well as optimization of individual VHH domains.

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。未满足的需求：基于骆驼科动物的抗体，具有上级抗原结合和物理活性， 化学性质（稳定性、亲水性等）没有充分发挥其潜力， 抗体有。这是因为骆驼免疫接种造成的后勤和财务障碍 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 动物产品：在这个SBIR项目中，我们建议在鼠宿主中开发一个遗传平台，用于 发现和开发含有骆驼科VHH的部分人源化杂合HCAb（及其产物） 域-在自然界中没有发现的组合。自20世纪90年代初被发现以来，HCAb已经产生了 由于其固有特性，对生物技术、诊断和治疗领域的兴趣不断增加， 适应性意义：除了小尺寸搭配坚固性和上级访问困难 表位，HCAb可以很容易地加工成，并利用作为，单域结合单位（VHH）， 保持它们对抗原的亲和力（与常规抗体相反）。创新：建议 携带预先工程化的羊驼/人免疫球蛋白基因座的靶向小鼠模型将增强 通过充当替代的杂合Ab宿主产生高亲和力HCAb。它将允许自然的，在体内 抗原特异性HCAb在小动物平台中的亲和力成熟，一个适合进一步遗传学研究的平台。 操纵它将使比自然骆驼宿主更大的队列规模成为可能，并简化HCAb 产生，从而为下游的HCAb和VHH结构域的开发提供了进一步的潜力。 应用.在我们的目标1中，我们专注于目标杂交羊驼/人的序列基因工程 胚胎干细胞中的免疫球蛋白位点。在目标2中，我们的重点是表征HCAbs的生成 源自新生成的杂交小鼠模型的免疫系统，并验证 我们的小动物平台。从我们的新型小鼠模型中获得的HCAbs将进行基本的 免疫学分析使我们能够表征抗体对模型的应答的库和效率， 抗原（I期）。如果成功，我们将利用新产生的动物模型生产HCAbs， 疾病相关的，困难的抗原和进展，以杂交瘤的发展，在进一步的合作（第二阶段）。 除此之外，我们的平台，基于其设计，还将允许进一步的基因修改发生 涉及来自其他物种的抗体结构域和/或进一步改变以增强HCAb多样性，以及 单个VHH结构域的优化。