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

这项研究的主要目标是开发一种在单抗生产过程中在线控制蛋白质糖基化的策略。PI将开发和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：在用中国仓鼠卵巢(CHO)细胞制造单抗时，如何实现对蛋白质糖基化的有效在线控制，从而确保单抗在体内具有可接受的生物活性？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，以将生物反应器的条件始终如一地保持在所需的设定点。任务2：先进的生物过程开发和优化：建立稳定的单抗生产CHO K1细胞系；开发用于量化宏观和微观异质性的糖基化分析；并确定稳健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：开发使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。单抗是一种对靶抗原表现出高度特异性的蛋白质，被用于治疗肿瘤学、器官移植、炎症性疾病等。目前有200多种单抗正在开发中，已成为生物制药行业增长最快的部门。与其他制造的产品一样，单抗只有在其产品质量属性(生物活性、效力、纯度等)时才具有治疗效果。位于特定值范围内。然而，满足这些通常严格的质量标准要求目前对制造商来说是一个重大挑战，因为MAB的制造过程非常复杂，而且没有很好地表征。此外，在目前的技术下，大多数质量控制分析必须离线和后期生产。尽管如此，保证所有药品的安全性和有效性的必要性促使FDA强烈建议制造商证明有能力在生产过程中在线确保产品质量。到目前为止，在MAB制造中还没有这样的在线质量控制技术。这项研究将在实验上证明糖基化的在线控制，并建立在实践中实现这一具有挑战性的目标的一般原则。PIs为这项研究设想了两种主要影响：(I)技术：对研究的总体成功至关重要的许多开创性技术的开发，特别是：细胞内分子过程的多尺度建模和控制；以及非线性半间歇生物过程的多速率、多回路控制系统的设计/实施。(Ii)实施：演示如何通过OPC软件将各种类型的测量系统(探头、分析仪和分析)与控制系统集成在一起--OPC软件是过程设备和控制系统之间通用连接的新兴行业标准。更广泛的影响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先者参与其中，并将结果传输给他们的客户。其次，开发的在线控制糖基化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实现质量控制的方式。第三，在为产生初步结果提供初始资金方面，FDA计划就特拉华大学的实验系统对检查员进行培训。第四，研究成果将被纳入过程动力学和控制课程，并通过出版物和向教育工作者和研究人员的演示广泛传播。最后，作为少数族裔，PI致力于招募代表性不足的团体进入化学工程学科，并应能够吸引少数族裔学生参与这一努力。目前正在参与该项目的研究生是女性，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.