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 用杂交瘤或原代B细胞体外筛选技术生产抗体 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量、效率和质量 抗体的筛选和制备方法。尽管这些产品具有大规模和高质量的要求 在工业中，选择用于生产的特定抗体的过程自动化仍然是一个未得到满足的需求。 CellRaft技术代表了一种新的成像、识别和分离单细胞和克隆的方法 殖民地。通过使用CellRaft AIR®系统对专有CellRaft阵列上的细胞进行成像，表型可以 在分离细胞和菌落用于下游繁殖之前，要详细地和随着时间的推移进行表征。在.期间 在第一阶段计划中，我们测试和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查平台的检测。简而言之， 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，用于种植和培养细胞。细胞的表型被监测在 具有CellRaft AIR系统成像能力的阵列。使用CellRaft细胞分析软件， 随着时间的推移，可以跟踪细胞并分析各种表型，包括荧光强度以及 向克隆克隆群体扩张。空气调节系统提供自动化、经济实惠、高效和坚固的 用于筛选单抗产生细胞的生产、亲和力和功能的平台 与世隔绝，因此只需要收获最有前途的候选人。在第二阶段，我们将改编 杂交瘤和Jurkat报告细胞系共培养，在第一阶段开发，能够筛选数百人 CellRaft-HTS阵列上数以千计的初级B细胞。我们将针对以下方面评估高吞吐量工作流 评估针对治疗相关抗原靶标的新型抗体的产生和功能。 为这一具有挑战性的工作流程提供自动化解决方案的当前技术无法与成本相媲美 节省，吞吐量，以及这里提出的详细的表型和功能表征。