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.

项目概要 通过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产品（包括研究）中最工业化的过程之一。 试剂、诊断测试和生物制药依赖于不同的通量、效率和质量 尽管这些抗体筛选和制造方法具有大规模和高质量的要求。 在工业领域，选择特定抗体进行生产的过程的自动化仍然是一个未得到满足的需求。 CellRaft 技术代表了一种成像、识别和分离单细胞和克隆细胞的新方法。 通过使用 CellRaft AIR® 系统对专有 CellRaft 阵列上的细胞进行成像，可以观察表型。 在分离细胞和集落进行下游繁殖之前，要详细地、随时间地进行表征。 在第一阶段计划中，我们测试并开发了新型报告细胞系、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的检测。 CellRaft 技术依赖于 CellRaft 阵列，该阵列包含数千个微孔，每个微孔具有一个 可释放的聚苯乙烯地板，用于对细胞进行表型监测。 具有 CellRaft AIR 系统成像功能的阵列，使用 CellRaft 细胞计数分析软件， 可以随着时间的推移跟踪细胞并分析各种表型，包括荧光强度以及 AIR 系统提供自动化、经济高效、高效且稳健的克隆克隆克隆扩展。 用于筛选单克隆抗体产生细胞的产量、亲和力和功能的平台 隔离，因此只有最有希望的候选者需要在第二阶段中被收获。 杂交瘤和 Jurkat 报告细胞系共培养在第一阶段开发，能够筛选数百个 我们将评估 CellRaft-HTS 阵列上数千个原代 B 细胞的高通量工作流程。 评估针对治疗相关抗原靶标的新型抗体的产生和功能。 当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、吞吐量以及此处提出的详细表型和功能表征。