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

摘要 单抗疗法现在占了大多数重磅炸弹药物的数量和广度 通过这种治疗方式解决的疾病的数量预计将在未来十年迅速增长。然而， 现有的发现技术迫使用户在性能和成本之间做出选择。有必要 高级发现工具，可在初始高吞吐量期间提供更好的性能功能性图景 放映。能够使抗体发现大众化的理想解决方案也将很容易获得，利用 制药、生物技术和研究实验室的现有设备。Partilion正在发展中， 商业化一种特殊的水凝胶微粒试剂(纳米瓶)，使超高通量成为可能 基于分泌抗体的功能特性的单个抗体分泌细胞的分类，均使用标准 实验室设备和随处可见的流式细胞仪。拓展一期工作并成功示范 在这个第二阶段的提案中，Partilion将从浆细胞中发现基于抗原特异性结合的抗体 开发纳米瓶工作流程，以实现单细胞功能分析(分泌抗体与细胞表面的结合 表达的靶和受体触发信号通路激活)为我们的最终用户。这些功能 可以通过最大限度地减少不起作用的 必须合成、导入细胞系、以大孔板格式生产和测试的序列 在下游。在这里，我们建议开发纳米瓶上的功能筛选分析，用于(I)与细胞表面的结合- 表达靶点和(Ii)受体激动性，并对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 诱饵工作流。使更多的公司和机构能够获得尖端药物发现能力 经济上也将推动罕见疾病治疗候选药物的进一步发展，并更加拥挤 “常见疾病”，这最终将为卫生系统节省费用，并为更多的人带来更好的结果 病人。

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.