The PROTEOMETER; A Continuous Upstream Process Monitoring Engine

Proteometer；一种连续的上游过程监控引擎 小灵通398具体研究计划 一、项目总结。 年，治疗性单抗(T-mAbbs)的全球市场规模为1050亿美元 2018年；临床诊断市场规模的12倍。除了对经济的影响外， 单抗已经成为医学界的一股主要力量。这项提案针对的是单抗 可以产生活性和免疫原性降低的蛋白形式变体，但不能 在生产过程中受到监控。FDA强调了检测和补救这一问题的必要性 问题自2004年以来一直没有成功1。Proteoform特定分析不适用于 发酵罐中蛋白质形式的连续监测。诺维莱特公司在此提出了一种新的 用于连续上游流程(CUSP)监控的分析引擎。 正在开发的CUSP Proteeter将有两个分析部门在 平行于从发酵罐中提取样品，分解单抗蛋白形式，每15- 30min，并评估mAb蛋白形式的异常产生。蛋白质形式比率的偏差将是 通过Proteeter检测到，潜在异常的信号将触发深度自下而上 分析。异常蛋白形式的嫌疑人将在70℃的固定化胰酶中消化 酶反应器，通过亲和选择将出水分解为糖肽和非糖肽 组分，以及以PTM特定方式检查的组分。携带PTM的多肽变体将 用多个柱子进行量化和识别。将有300多个无人参与的分析 在为期两周的单抗生产活动中运行。 CUSP Proteeter数据管理系统将区块链编码mAb结构 和关系源、时间、协议、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 制造业，以及潜在的异常情况。在迅速确认了一种 它可以由FDA“实时”发布。此外，这些数字历史可以是 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的要求改进制造。

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.