The PROTEOMETER; A Continuous Upstream Process Monitoring Engine

蛋白质计；

• 批准号: 10005833
• 负责人: Jinhee Kim
• 金额: $ 68.97万
• 依托单位: NOVILYTIC, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-02 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10005833
• 关键词: Address; Affinity; Antibodies; Biological Assay; Biological Products; Cells; Chromatography; Code; Collaborations; Communication; Computer software; Data; Data Management Resources; Development; Equipment; Failure; Fermentation; Glycopeptides; Goals; Health; High Pressure Liquid Chromatography; Hour; Immobilized Enzymes; Industry; Island; Libraries; Manufacturer Name; Medicine; Monitor; Monoclonal Antibodies; Operating System; Patients; Peptides; Performance; Phase; Process; Production; Proteins; Protocols documentation; Recording of previous events; Research; Running; Safety; Sampling; Signal Transduction; Societies; Source; Stress; Structure; System; Testing; Therapeutic Monoclonal Antibodies; Thinness; Time; Trypsin; Validation; Variant; blockchain; clinical diagnostics; cost; digital; economic impact; encryption; enzyme reactor; glycosylation; immunogenicity; improved; monoclonal antibody production; protein aggregation; remediation; research and development; software development

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.

蛋白质计；连续的上游过程监控引擎 PHS 398 具体研究计划 一、项目概要。 治疗性单克隆抗体 (T-mAb) 的全球市场规模为 1050 亿美元 2018；是临床诊断市场规模的十二倍。除了经济影响之外， 单克隆抗体已成为医学领域的一支重要力量。该提案解决了以下事实：mAb 可以产生活性和免疫原性降低的蛋白质变体，但不能 生产过程中进行监控。 FDA 强调需要检测和补救这一问题 自 2004 年以来一直存在这个问题，但没有成功1。 Proteoform 特定分析不适用于 连续监测发酵罐中的蛋白质形式。 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 并行从发酵罐中提取样品，解析 mAb 蛋白质形式，每 15- 对其进行定量 30 分钟，并评估异常 mAb 蛋白形式的产生。蛋白质型比率的偏差将为 由 Proteometer 感测到，潜在异常的信号将触发深入的自下而上 分析。异常的蛋白质形式可疑物将在固定化的胰蛋白酶中于 70 oC 下消化 酶反应器，流出物通过亲和选择分为糖肽和非糖肽 分数，以及以 PTM 特定方式检查的分数。带有 PTM 的肽变体将 通过多列进行量化和识别。将进行 300 多项无人值守分析 在为期两周的单克隆抗体生产活动期间运行。 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确定数据真实性和单克隆抗体质量 制造和潜在的异常。迅速确认了产品的安全性和质量 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 的要求改进制造。