GOALI: Multi-scale Modeling and Advanced Control of Glycosylation in Monoclonal Antibody Production

本研究的主要目的是开发一种在单克隆抗体生产过程中在线控制蛋白糖基化的策略。pi将开发并实验验证一种新型的多回路在线控制系统，该系统将多尺度模型与综合多速率测量系统集成在一起。需要回答的具体问题是：当使用中国仓鼠卵巢（CHO）细胞制造单克隆抗体（MAb）时，如何实现有效的蛋白质糖基化在线控制，从而确保单克隆抗体在体内可接受的生物活性？将执行的具体任务是：任务1：内环控制系统开发：开发和实施多变量内环控制系统，以保持生物反应器条件始终保持在所需的设定点。任务2：先进的生物工艺开发和优化：开发稳定的产生单克隆抗体的CHO K1细胞系；发展糖基化分析，定量分析宏观和微观异质性；并确定稳定获得质量的最佳条件，即所需的糖基化模式和活性。任务3：外环控制系统开发；整体系统集成/实施：制定采用糖基化测量和糖基化状态估计的外环控制方案，将其与内环集成，并实施完整的多环控制方案，以演示糖基化的有效在线控制。单克隆抗体（mab）是一种对靶抗原具有高特异性的蛋白质，用于肿瘤、器官移植、炎症性疾病等治疗。随着200多种单克隆抗体的开发，单克隆抗体已成为生物制药行业增长最快的部门。与其他制成品一样，单克隆抗体只有在其产品质量属性（生物活性、效力、纯度等）处于特定值范围内时才具有治疗效果。然而，满足这些通常严格的质量标准要求目前是制造商面临的主要挑战，因为单克隆抗体的制造过程非常复杂，并且没有很好地表征。此外，根据目前的技术，大多数质量控制分析必须在离线和生产后进行。然而，保证所有药品的安全性和有效性的必要性促使FDA强烈建议制造商在生产过程中证明有能力在线确保产品质量。到目前为止，在MAb制造中还没有这样的在线质量控制技术。本研究将通过实验证明糖基化的在线控制，并为在实践中实现这一具有挑战性的目标建立一般原则。pi设想了这项研究的两种主要影响：(i)技术：许多开创性技术的发展对研究的整体成功至关重要，特别是：细胞中分子过程的多尺度建模和控制；以及非线性半批量生物过程的多速率、多回路控制系统的设计/实现。（ii）实施：演示如何通过OPC软件将各种类型的测量系统（探针、分析仪和分析）与控制系统集成？用于过程设备和控制系统之间通用连接的新兴工业标准。本研究的结果将对工业实践产生直接影响。首先，生物过程监测和控制系统硬件的行业领导者将参与其中，并将结果传递给他们的客户。其次，为在线控制糖基化而开发的策略可以应用于任何治疗性蛋白质产品的其他质量属性。因此，如果成功，这项工作可能会彻底改变生物制药行业如何实现质量控制。第三，在为产生初步结果提供初始资金的同时，FDA计划培训特拉华大学实验系统的检查员。第四，将研究成果纳入过程动力学与控制课程，并通过出版物和报告向教育工作者和研究人员广泛传播。最后，PI本身作为少数民族，致力于在化学工程学科中招募代表性不足的群体，并且应该能够吸引少数民族学生参与这一努力。目前从事该项目的研究生是一名女性，她产生了初步结果。

1034213Ogunnaike The primary goal of this research is to develop a strategy for on-line control of protein glycosylation during monoclonal antibody production. The PIs will develop and validate experimentally, a novel multi-loop, on-line control system incorporating a multi-scale model integrated with a comprehensive multi-rate measurement system. The specific question to be answered is: When manufacturing monoclonal antibodies (MAb) using Chinese Hamster Ovary (CHO) cells, how does one achieve effective on-line control of protein glycosylation and hence assure acceptable MAb bioactivity in vivo? The specific tasks that will be performed are: Task 1: Inner-loop Control System Development: develop and implement a multivariable inner-loop control system to maintain bioreactor conditions consistently at desired set-points. Task 2: Advanced Bioprocess Development and Optimization: develop a stable MAb-producing CHO K1 cell line; develop glycosylation assays for quantifying both macro- and micro-heterogeneity; and determine the optimal conditions for robust attainment of quality i.e., desired glycosylation pattern and activity. Task 3: Outer-loop Control System Development; Overall System Integration/Implementation: develop an outer-loop control scheme which employs glycosylation measurements and state estimates of glycosylation, integrate this with the inner loop, and implement the complete multi-loop control scheme to demonstrate effective on-line control of glycosylation. Monoclonal antibodies (MAbs), proteins that exhibit high specificity for a target antigen, are used therapeutically in oncology, organ transplantation, inflammatory disease, etc. With more than 200 in development pipelines, MAbs have become the fastest growing sector of the biopharmaceutical industry. As with other manufactured products, MAbs are therapeutically effective only when their product quality attributes (bioactivity, potency, purity, etc.) lie within a specific range of values. However, meeting these often stringent quality criteria requirements is currently a major challenge for manufacturers because the MAb manufacturing process is very complex and not well-characterized. In addition, with current technology, the majority of quality control assays must be performed off-line and post-production. Nevertheless, the necessity to guarantee the safety and effectiveness of all pharmaceutical products has prompted the FDA to recommend strongly that manufacturers demonstrate the ability to ensure product quality online during production. To date, no such technique for online quality control in MAb manufacturing exists. Intellectual Merit This research will demonstrate experimentally, on-line control of glycosylation, and establish general principles for achieving such a challenging objective in practice. The PIs envision two kinds of primary impact for this research: (i) Technical: the development of many pioneering techniques central to the overall success of the research, specifically: multi-scale modeling and control of molecular processes in the cell; and the design/ implementation of multi-rate, multi-loop control systems for non-linear semi-batch bioprocesses. (ii) Implementational: a demonstration of how to integrate various categories of measurement systems (probes, analyzers, and assays) with control systems via OPC software?the emerging industry standard for universal connectivity between process equipment and control systems. Broader Impact The outcomes of this research will have a direct impact on industrial practice. First, industry leaders in bioprocess monitoring and control systems hardware are involved, and will transmit the results to their customers. Second, the strategy developed for on-line control of glycosylation can be applied to other quality attributes in any therapeutic protein product. Thus, if successful, this work could potentially revolutionize how quality control is achieved in the biopharmaceutical industry. Third, in providing initial funding for generating preliminary results, the FDA plans to train inspectors on the University of Delaware experimental system. Fourth, the research results will be incorporated into the process dynamics and control course, and widely disseminated through publications and presentations to educators and researchers. Finally, the PI, as a minority himself, is committed to recruiting under-represented groups into the chemical engineering discipline in general and should be able to attract minority students to participate in this effort. The graduate student currently working on the project, who generated the preliminary results, is female.