The PROTEOMETER; A Continuous Upstream Process Monitoring Engine

PROTEOMETER;连续上游过程监测引擎 PHS 398特定研究报告 I.项目摘要。 2010年，全球治疗性单克隆抗体（T-mAbs）市场为1050亿美元。 2018年;临床诊断市场规模的12倍。除了经济影响之外， 单克隆抗体已成为医学中的主要力量。该提案解决了mAb 可以产生活性和免疫原性降低的蛋白形式变体，但不 在生产过程中进行监控。FDA强调有必要检测和补救这一点 自2004年以来一直没有成功。Proteoform特定分析不适用于 连续监测发酵罐中的蛋白质型。Novilytic在此提出了一种新的 用于连续上游过程（CUSP）监控的分析引擎。 正在开发的CUSP Proteometer将有两个分析部门， 平行于从发酵罐中提取样品，解析mAb蛋白质型，每15分钟定量一次。 30分钟，并评估异常的mAb蛋白质型产生。蛋白质形式比率的偏差将为 通过蛋白质仪的感知，潜在异常的信号将触发深入的自下而上 分析。将异常蛋白形式可疑物在70 oC的固定化 酶反应器，流出物通过亲和选择分成糖肽和非糖肽 级分，以及以PTM特定方式检查的级分。携带PTM的肽变体将 可以用多个列进行量化和标识。将进行300多项无人值守分析， 在两周的mAb生产活动期间运行。 CUSP Proteometer数据管理系统将区块链编码mAb结构 以及关系源、时间、协议和设备性能块中的环境数据; 使联邦监管机构能够在整个过程中建立数据真实性、mAb质量 制造和潜在的异常。在迅速确认了安全和质量的一个 mAb批次可以由FDA“实时”发布。此外，这些数字历史可以 内置于库中，用于识别和修复流程弱点，增强流程 分析，并按照FDA的要求改进生产。

The PROTEOMETER; A Continuous Upstream Process Monitoring Engine PHS 398 SPECIFIC RESEARCH PLAN I. PROJECT SUMMARY. The global market for therapeutic monoclonal antibodies (T-mAbs) was $105 billion in 2018; twelve times the size of the clinical diagnostics market. Beyond their economic impact, mAbs have become a major force in medicine. This proposal addresses the fact that mAb proteoform variants of diminished activity and immunogenicity can be generated, but not monitored during production. The FDA has stressed the need to detect and remediated this problem since 2004 without success1. Proteoform specific analytics are not available for continuous monitoring of proteoforms in a fermentor. Novilytic is proposing herein a new analytical engine for continuous upstream process (CUSP) monitoring. The CUSP Proteometer being developed will have two analytical sectors working in parallel to extract samples from a fermentor, resolve mAb proteoforms, quantify them every 15- 30 min, and assess aberrant mAb proteoform production. Deviation in proteoform ratio will be sensed by the Proteometer, signaling of a potential aberrant will trigger in-depth bottom-up analyses. Aberrant proteoform suspects will be trypsin digested at 70 oC in an immobilized enzyme reactor, the effluent split by affinity selection into glycopeptide and non-glycopeptide fractions, and the fractions examined in PTM specific ways. PTM bearing peptide variants will be quantified and identified with multiple columns. More than 300 unattended analyses will be run during a two week mAb production campaign. The CUSP Proteometer data management system will blockchain code mAb structural and environmental data in relational source, time, protocol, and equipment performance blocks; enabling federal regulatory agencies to establish data authenticity, mAb quality throughout manufacturing, and potential aberrants. Having rapidly confirmed the safety and quality of a mAb lot it can be released by the FDA in “real-time”. Additionally, these digital histories can be built into a library and used to identify and remediate process weaknesses, enhance process analytics, and improve manufacturing as mandated by the FDA.