Rapid identification and selection of functional antigen-specific monoclonalantibodies by FcGR-enabled screening on CellRaft Arrays in the CellRaft AIRSystem

通过在 CellRaft AIRSystem 中的 CellRaft 阵列上进行 FcGR 筛选，快速鉴定和选择功能性抗原特异性单克隆抗体

• 批准号: 10698784
• 负责人: Jessica Hartman
• 金额: $ 83.81万
• 依托单位: CELL MICROSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10698784
• 关键词: Affinity; Animals; Antibodies; Antibody Formation; Antibody-Producing Cells; Antigen Targeting; Antigens; Automation; B-Lymphocytes; Binding; Binding Proteins; Biological Assay; Biological Products; Biological Sciences; Cell Culture Techniques; Cell Line; Cell Separation; Cells; Clinical; Cloning; Coculture Techniques; Computer software; Cost Savings; Cytometry; Data; Development; Diagnostic tests; ERBB2 gene; Egg Proteins; Epidermal Growth Factor Receptor; Floor; Fluorescence; GPC3 gene; Generations; Goals; Growth; Harvest; Hybridomas; Image; Immune; Immunize; In Vitro; Individual; Industrialization; Industry; Laboratories; Libraries; Licensing; Marketing; Medicine; Methods; Modern Medicine; Monitor; Monoclonal Antibodies; Mus; Phase; Phenotype; Polystyrenes; Process; Production; Proteins; Reagent; Recovery; Reporter; Research; Screening procedure; Series; Signal Transduction; Site; Sorting; Splenocyte; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Monoclonal Antibodies; Time; Tumor Antigens; Universities; Vaccinated; Vaccination; Validation; Work; antibody libraries; cellular imaging; commercialization; cost; cost effective; cost efficient; high throughput screening; imaging capabilities; improved; in vitro testing; in vivo; lead candidate; manufacture; novel; nuclear factors of activated T-cells; phase 1 study; phase 2 study; programs; screening; success; tool

Project Summary The production of antibodies using hybridoma or primary B-cells with in vitro screening technologies represents one of the most industrialized processes in contemporary life science. Products including research reagents, diagnostic tests and biopharmaceuticals rely on the throughput, efficiency and quality of different antibody screening and manufacturing methods. Despite the large-scale and high-quality requirements of these industries, automation of the process for selecting specific antibodies for manufacturing remains an unmet need. The CellRaft Technology represents a novel means of imaging, identifying, and isolating single cells and clonal colonies. By imaging cells on the proprietary CellRaft Array using the CellRaft AIR® System, phenotypes can be characterized in detail and over time, prior to isolating cells and colonies for downstream propagation. During the Phase I program, we tested and developed novel reporter cell lines, software, and cell-based co-culture assays that leveraged our CellRaft AIR System as an automated antibody screening platform. Briefly, the CellRaft Technology relies on the CellRaft Array, which contains thousands of microwells, each featuring a releasable polystyrene floor where cells are seeded and cultured. Cells are phenotypically monitored on the array with the imaging capabilities of the CellRaft AIR System. Using the CellRaft Cytometry analytical software, cells can be tracked over time and analyzed for various phenotypes, including fluorescence intensity, as well as expansion into clonal colonies. The AIR System provides an automated, cost-effective, efficient, and robust platform for screening the production, affinity and functionality of monoclonal antibody producing cells prior to isolation so only the most promising candidates need to be harvested. During Phase II, we will adapt the hybridoma and Jurkat reporter cell line co-culture that was developed in Phase I to be able to screen hundreds of thousands of primary B cells on the CellRaft-HTS Array. We will evaluate a high throughput workflow for assessing production and functionality of novel antibodies against therapeutically relevant antigen targets. Current technologies offering automated solutions to this challenging workflow are incapable of rivaling the cost savings, throughput, and the detailed phenotypic and functional characterization proposed here.

项目摘要 利用体外筛选技术使用杂交瘤或原代B细胞生产抗体 代表了当代生命科学中最工业化的过程之一。产品包括研究 试剂、诊断测试和生物制药依赖于不同的生产能力、效率和质量。 抗体筛选和制造方法。尽管大规模和高质量的要求，这些 在工业上，选择用于制造的特异性抗体的过程的自动化仍然是未满足的需求。 CellRaft技术代表了一种成像、鉴定和分离单细胞和克隆的新方法。 殖民地通过使用CellRaftAIR ®系统在专有的CellRaftArray上对细胞进行成像， 在分离细胞和集落用于下游增殖之前，应详细表征并随时间推移。期间 在I期项目中，我们测试并开发了新的报告细胞系、软件和基于细胞的共培养 利用我们的CellRaft AIR系统作为自动化抗体筛选平台进行检测。采用离体垂体碎片 CellRaft技术依赖于CellRaft阵列，该阵列包含数千个微孔，每个微孔具有 可释放的聚苯乙烯地板，其中细胞被接种和培养。细胞表型监测在 阵列与CellRaft AIR系统的成像能力。使用CellRaft Cytometry分析软件， 可以随时间跟踪细胞并分析各种表型，包括荧光强度，以及 扩展成克隆菌落。AIR系统提供了一个自动化的，具有成本效益的，高效的，强大的 用于筛选单克隆抗体产生细胞的产生、亲和力和功能性的平台， 隔离，因此只有最有希望的候选者需要被收获。在第二阶段，我们将调整 杂交瘤和Jurkat报告细胞系共培养，在I期开发，能够筛选数百个 CellRaft-HTS阵列上的数千个主要B细胞。我们将评估一个高通量的工作流程， 评估针对治疗相关抗原靶标的新型抗体的产生和功能性。 目前为这一具有挑战性的工作流程提供自动化解决方案的技术无法与成本相媲美 节省、通量以及这里提出的详细的表型和功能表征。