Linking antibody sequences to function at the single-cell level using nanovial technology

使用纳米瓶技术连接抗体序列以在单细胞水平发挥作用

• 批准号: 10697372
• 负责人: Joseph de Rutte
• 金额: $ 76.76万
• 依托单位: PARTILLION BIOSCIENCE CORPORATION
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-16 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10697372
• 关键词: Adoption; Affinity; Agonist; Antibodies; Antibody Diversity; Antibody Repertoire; Antigen Targeting; Antigens; Autoimmune Diseases; B-Cell Antigen Receptor; Bacteriophages; Benchmarking; Binding; Binding Proteins; Biological Assay; Biotechnology; COVID-19; Capital; Cell Line; Cell Separation; Cell membrane; Cell surface; Cells; Cellular Assay; Chromium; Cost Savings; Crowding; Data; Democracy; Development; Disease; Equipment; Expenditure; Genomics; Health system; Hybridomas; Hydrogels; Immunization; Immunoglobulin-Secreting Cells; Immunologics; Incubated; Interview; Label; Laboratory Research; Light; Link; MS4A1 gene; Malignant Neoplasms; Marketing; Membrane; Methodology; Microfluidics; Modality; Modeling; Monoclonal Antibody Therapy; Outcome; Patients; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Plasma Cells; Process; Property; Rare Diseases; Reagent; Receptor Signaling; Reporter; Research; Running; Scientist; Side; Signal Pathway; Signal Transduction; Small Business Innovation Research Grant; Sorting; Surface Antigens; System; TNFRSF1A gene; Technology; Testing; Therapeutic; Tissues; Training; Tube; Virus Diseases; Work; Yeasts; antagonist; commercialization; cost; drug discovery; falls; fluorescence activated cell sorter device; high throughput screening; improved; instrument; laboratory equipment; particle; receptor; screening; single cell sequencing; therapeutic candidate; therapeutic development; therapeutic target; tool

ABSTRACT Monoclonal antibody therapies now account for the majority of blockbuster drugs and the number and breadth of diseases tackled by this therapeutic modality are expected to rapidly grow in the coming decade. However, existing discovery technologies force users to choose between performance and cost. There is a need for advanced discovery tools that provide a better functional picture of performance during initial high-throughput screening. An ideal solution that can democratize antibody discovery would also be easily accessible, leveraging existing equipment in pharmaceutical, biotech, and research laboratories. Partillion is developing and commercializing a specialized hydrogel microparticle reagent (“nanovials”) that enables ultra high-throughput sorting of single antibody secreting cells based on functional properties of secreted antibodies, all using standard lab equipment and widely-available flow cytometers. Expanding on Phase I work and successful demonstration of antibody discovery from plasma cells based on antigen-specific binding, in this Phase II proposal Partillion will develop nanovial workflows to enable single-cell functional assays (binding of secreted antibodies to cell-surface expressed targets and receptors that triggers signaling pathway activation) for our end users. These capabilities can provide dramatic improvements in discovery workflows by minimizing the number of non-functional sequences that have to be synthesized, introduced into cell lines, produced, and tested in large well-plate formats downstream. Here we propose to develop on-nanovial functional screening assays for (i) binding to cell-surface- expressed targets and (ii) receptor agonism, and benchmark the antibody sequences discovered with these approaches against sequences recovered using standard hybridoma, and direct B cell Receptor (BCR) antigen baiting workflows. Enabling more companies and institutes to access cutting-edge drug discovery capabilities economically will also drive further development of therapeutic candidates for rare diseases and more crowded “common diseases”, which ultimately will yield cost savings to the health systems and better outcomes for more patients.

抽象的 单克隆抗体疗法目前占据重磅药物的大部分，且数量和广度 通过这种治疗方式治疗的疾病预计在未来十年将迅速增长。 现有的发现技术迫使用户在性能和成本之间做出选择。 先进的发现工具，可在初始高通量期间提供更好的性能功能图 利用筛选，可以轻松实现抗体发现民主化的理想解决方案。 Partillion 正在开发和研究制药、生物技术和研究实验室的现有设备。 将一种特殊的水凝胶微粒试剂（“nanovials”）商业化，以实现超高通量 根据分泌抗体的功能特性对单个抗体分泌细胞进行分类，全部使用标准 实验室设备和广泛使用的流式细胞仪扩大了第一阶段的工作并成功示范。 在此 II 期提案中，Partillion 将基于抗原特异性结合从浆细胞中发现抗体 开发纳米瓶工作流程以实现单细胞功能测定（分泌的抗体与细胞表面的结合） 为我们的最终用户表达触发信号通路激活的靶标和受体。 可以通过最大限度地减少非功能性的数量来显着改进发现工作流程 必须在大型孔板中合成、引入细胞系、生产和测试的序列 在这里，我们建议开发纳米功能筛选试验，用于（i）与细胞表面的结合。 表达的目标和（ii）受体激动，并对用这些发现的抗体序列进行基准测试 针对使用标准杂交瘤和直接 B 细胞受体 (BCR) 抗原恢复的序列的方法 使更多的公司和机构能够获得尖端的药物发现能力。 经济上也将进一步推动罕见疾病和更拥挤的治疗候选药物的开发 “常见疾病”，最终将为卫生系统节省成本，并为更多人带来更好的结果 患者。