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

项目摘要 转基因小鼠模型已被证明是产生抗体相关的必不可少的 生物药物(生物制品)。到目前为止，大多数生物制剂来自小鼠模型和小动物 模型不仅用于生成抗体，还用作进一步优化和测试的平台 生物制品中的。基于骆驼的抗体，具有良好的抗原结合和物理化学特性 性能(稳定性、亲水性等)还没有充分发挥他们的潜力，在同样的程度上 传统的抗体有。这是建立在对骆驼的后勤和财政障碍免疫上的。 准备生产单抗重链抗体(HCAb)，以及体外技术不能 全面概括了对极其多样化、高亲和力的粘合剂的特殊自然选择 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基因平台 发现和开发部分人源化的杂合HCAbs(及其产品)。自从他们在 20世纪90年代初，HCAbs在生物技术、诊断和治疗领域引起了越来越多的兴趣 由于其固有的特性和适应性。除了体积小，坚固耐用，性能出众 获得困难的表位，HCAb可以很容易地加工成单域结合单位并用作单域结合单位 (VHH)，同时保持其对抗原的亲和力(与传统抗体相反)。建议数 携带工程免疫球蛋白基因的靶向小鼠模型将增强高表达 通过作为另一种杂交抗体宿主来亲和HCAbs。它将允许自然的、体内的亲和力成熟 抗原特异性HCAb在一个小动物平台上，一个服从于进一步的基因操作。它将使 比天然骆驼宿主更大的队列大小，并简化了HCAb的生成，从而提供了进一步的 为下游应用开发HCAb和VHH结构域的潜力。在我们的目标1中，我们专注于 关于通过增加更多的骆驼VHH来磨练和鉴定我们的杂交骆驼免疫球蛋白基因座 将改良的人VHS导入该基因座，同时评估B细胞发育和抗体亲和力 和多样性。在目标2中，我们的重点是通过以下方式对抗体应答的指标和效率进行基准测试 通过使用与疾病相关的困难抗原和对杂交瘤的有希望的打击进展来竞争技术 开发和规模化抗体生产。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并建立 制药公司支持发现高价值靶标的新型抗体。

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.