摘 要
生物制药作为现代医药工业的重要组成部分，其产品质量和生产效率在很大程度上依赖于中下游分离纯化工艺的开发与优化。本研究旨在针对目标生物分子的理化特性和生物学功能，设计并验证高效、稳定的分离纯化工艺流程。通过结合层析技术、膜过滤技术和结晶等方法，构建了多步联用的分离纯化体系，并引入新型填料和智能化控制策略以提升工艺性能。实验中采用模型蛋白和重组表达的目标蛋白进行工艺开发，通过对关键工艺参数（如pH值、离子强度、流速等）的系统性优化，显著提高了目标产物的回收率和纯度。同时，基于质量源于设计（QbD）理念，建立了风险评估模型和工艺验证方案，确保工艺的稳健性和可重复性。研究结果表明，所开发的工艺能够实现目标蛋白的高纯度分离（纯度>98%）和高回收率（回收率>85%），且具有良好的放大适应性。此外，本研究创新性地引入了在线监测和反馈控制系统，实现了对关键质量属性（CQA）的实时监控，为工艺的自动化和智能化奠定了基础。总体而言，本研究不仅为生物制药领域提供了高效的分离纯化解决方案，还为类似工艺的开发与验证提供了有价值的参考框架，具有重要的理论意义和实际应用价值。
关键词：生物制药；分离纯化工艺；层析技术；质量源于设计；在线监测
Abstract
Biopharmaceuticals, as a critical component of modern pharmaceutical industries, largely depend on the development and optimization of downstream separation and purification processes for product quality and production efficiency. This study focuses on designing and validating efficient and robust separation and purification workflows tailored to the physicochemical properties and biological functions of target biomolecules. By integrating chromatographic techniques, membrane filtration, and crystallization methods, a multi-step separation and purification system was constructed, with novel packing materials and intelligent control strategies introduced to enhance process performance. Model proteins and recombinantly expressed target proteins were utilized in the experimental process, and systematic optimization of key process parameters, such as pH, ionic strength, and flow rate, significantly improved the recovery and purity of the target products. Furthermore, based on the Quality by Design (QbD) concept, a risk assessment model and process validation scheme were established to ensure process robustness and reproducibility. The results demonstrated that the developed process achieved high-purity separation (purity > 98%) and high recovery (recovery > 85%) of the target protein, with excellent scalability. Additionally, this study innovatively incorporated online monitoring and feedback control systems to enable real-time surveillance of Critical Quality Attributes (CQAs), laying the foundation for the automation and intelligence of the process. Overall, this research not only provides an efficient separation and purification solution for the biopharmaceutical field but also offers a valuable reference fr amework for the development and validation of similar processes, highlighting significant theoretical implications and practical application value.
Keywords: Biopharmaceutical; Separation Purification Process; Chromatography Technology; Quality By Design; Online Monitoring
目 录
1绪论 1
1.1生物制药中下游工艺的研究背景 1
1.2分离纯化工艺开发的意义与价值 1
1.3国内外研究现状与发展趋势 1
1.4本文研究方法与技术路线 2
2分离纯化工艺的关键技术分析 2
2.1生物分子分离的基本原理 2
2.2色谱技术在纯化中的应用 3
2.3膜分离技术的优化策略 3
2.4工艺参数对分离效率的影响 4
2.5关键技术的选择与评价 4
3分离纯化工艺的开发流程 5
3.1工艺开发的目标与要求 5
3.2原料处理与预分离技术 5
3.3纯化步骤的设计与优化 6
3.4工艺放大中的挑战与解决方案 6
3.5开发流程的验证与改进 7
4分离纯化工艺的验证与质量控制 7
4.1验证体系的建立与实施 7
4.2工艺稳定性测试与评估 8
4.3产品质量标准与检测方法 8
4.4风险评估与控制措施 9
4.5验证结果的分析与总结 9
结论 11
参考文献 12
致 谢 13
