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Novel Process Analytic Technology for Continuous Bioprocessses

用于连续生物过程的新型过程分析技术

基本信息

批准号：
10238815
负责人：
JONGYOON HAN
金额：
$ 100万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10238815
关键词：
Novel Process Analytic Technology Continuous

项目摘要

Abstract This proposal aims to demonstrate how real-time measurement of protein product quality attributes and rapid assessment of adventitious agent contamination can be used for manufacturers’ risk-based decision making in continuous protein production. While Quality by Design has been a successful approach for maintaining product quality, its adoption is a byproduct of difficulties in measuring product quality in real time. We will experimentally demonstrate two novel platforms for in-line protein quality assessment (1) a micro/nanofluidic platform for assessing high- and low-molecular weight protein impurities, protein structure and binding, and glycosylation and (2) swept source Raman (SS-Raman) – using a novel approach of a tunable laser and low-cost, fixed-wavelength detector instead of a traditional, expensive spectrometer or interferometer – to measure process parameters and product quality at all points of the process. We have previously demonstrated the use of micro- and nano-fluidic assays to measure protein quality attributes off-line, constructed an automated sampling system to feed bioreactor supernatant to the micro/nanofluidic assays, and used Raman spectroscopy to identify and quantify therapeutic proteins dissolved in buffer solutions at concentrations found in drug product and at volumes equivalent to or less than one dose. This work will be the first demonstration of these technologies for real-time protein quality assessment. Additionally, this work will develop novel approaches for rapid adventitious agent detection and demonstrate their use in CHO cell culture. We will demonstrate the use of Raman spectroscopy to obtain metabolic fingerprints (intracellular metabolites) and footprints (extracellular metabolites) of protein producing CHO cells when challenged by various pathogens and classify these fingerprints and footprints as either normal or anomalous, indicating the presence of a potential infection. We will also evaluate rapid Next Generation Sequencing and bioinformatic approaches for their suitability to rapidly detect virus contaminations of continuous CHO cell culture. Finally, this proposal will investigate the impact of integrating these novel technologies into a continuous monoclonal antibody manufacturing process. At its completion this work will improve (a) product and process knowledge by demonstrating the use of real-time protein attribute measurements (b) process control by enabling direct measurement of critical quality attributes during manufacturing and feedback or feedforward control strategies to be tested, and (c) safety through development of rapid adventitious agent assays to more rapidly assure sterility of biopharmaceuticals which can improve patient safety, assure supply of medicines, and reduce business risk.

摘要本提案旨在演示如何实时测量蛋白质产品质量外源因子污染的属性和快速评估可用于制造商在连续蛋白质生产中基于风险的决策。质量由设计一直是保持产品质量的成功方法，它的采用是难以真实的实时测量产品质量的副产品。我们将实验性地展示了两种用于在线蛋白质质量评估的新平台（1）微/纳米流体用于评估高分子量和低分子量蛋白质杂质、蛋白质结构和结合和糖基化以及（2）扫频源拉曼（SS-拉曼）-使用新方法可调谐激光器和低成本的固定波长探测器，而不是传统的昂贵的光谱仪或干涉仪-测量所有点的工艺参数和产品质量的过程。我们以前已经证明了使用微流体和纳米流体测定，离线检测蛋白质质量属性，构建自动化进样系统，生物反应器上清液的微/纳米流体测定，并使用拉曼光谱法来鉴定并以药物中发现的浓度定量溶解在缓冲溶液中的治疗性蛋白质，产品和体积等于或小于一个剂量。这项工作将是第一个这些技术用于实时蛋白质质量评估的示范。而且这个工作将开发新的方法，快速检测外源因子，并证明其用于CHO细胞培养。我们将演示使用拉曼光谱来获得代谢产物蛋白质指纹（细胞内代谢物）和足迹（细胞外代谢物）当受到各种病原体攻击时产生CHO细胞，并对这些指纹进行分类，足迹正常或异常，表明存在潜在的感染。我们将我还评估快速下一代测序和生物信息学方法的适用性以快速检测连续CHO细胞培养物的病毒污染。最后，该提案将研究将这些新技术整合到连续单克隆抗体中的影响，抗体制造过程。这项工作完成后将改进（a）产品和工艺通过演示实时蛋白质属性测量（B）过程的使用获得知识通过在生产过程中直接测量关键质量属性进行控制，反馈或前馈控制策略进行测试，以及（c）通过开发快速外源因子测定，以更快地确保生物药物的无菌性，提高患者安全性，确保药品供应，降低商业风险。