摘 要
重组蛋白药物在现代生物医药领域中占据重要地位，其高效表达和质量控制依赖于合适的表达系统选择与优化策略。本研究以提高重组蛋白药物的产量和活性为目标，系统分析了不同表达系统的优劣，并提出了针对性的优化方案。研究首先回顾了常用的原核和真核表达系统的特点及其适用范围，包括大肠杆菌、酵母、昆虫细胞及哺乳动物细胞等平台，明确了各系统在蛋白折叠、翻译后修饰和稳定性方面的差异。在此基础上，通过实验验证了多种优化手段的有效性，例如启动子调控、信号肽设计、共表达分子伴侣以及培养条件改进等技术。结果表明，针对目标蛋白特性进行个性化表达系统设计能够显著提升蛋白表达水平和功能属性。此外，本研究创新性地引入了计算建模方法预测蛋白表达效率，并结合高通量筛选技术加速了优化流程。最终，该策略成功应用于多个重组蛋白药物的开发实例中，验证了其普适性和实用性。综上所述，本研究不仅为重组蛋白药物的表达系统选择提供了理论依据，还通过多维度优化显著提升了生产效率，为相关领域的进一步发展奠定了坚实基础。
关键词：重组蛋白药物；表达系统优化；翻译后修饰；计算建模；高通量筛选
Abstract
Recombinant protein drugs play a crucial role in modern biopharmaceuticals, with their efficient ex pression and quality control relying heavily on the selection and optimization of appropriate ex pression systems. This study aimed to enhance the yield and activity of recombinant protein drugs by systematically analyzing the advantages and disadvantages of various ex pression systems and proposing targeted optimization strategies. It began with a review of the characteristics and applicable scopes of commonly used prokaryotic and eukaryotic ex pression systems, including Escherichia coli, yeast, insect cells, and mammalian cells, clarifying the differences among these systems in terms of protein folding, post-translational modifications, and stability. Based on this analysis, the effectiveness of multiple optimization approaches was experimentally validated, such as promoter regulation, signal peptide design, co-ex pression of molecular chaperones, and improvement of culture conditions. The results demonstrated that personalized ex pression system design tailored to the properties of target proteins could significantly increase protein ex pression levels and functional attributes. Additionally, this study innovatively introduced computational modeling methods to predict protein ex pression efficiency and combined them with high-throughput screening technologies to expedite the optimization process. Ultimately, this strategy was successfully applied to several case studies of recombinant protein drug development, confirming its universality and practicality. In summary, this study not only provided theoretical evidence for the selection of ex pression systems for recombinant protein drugs but also substantially improved production efficiency through multi-dimensional optimization, laying a solid foundation for further advancements in related fields.
Keywords: Recombinant Protein Drugs; ex pression System Optimization; Post-Translational Modification; Computational Modeling; High-Throughput Screening
目 录
1绪论 1
1.1重组蛋白药物表达系统的研究背景 1
1.2选择与优化策略的意义分析 1
1.3国内外研究现状综述 1
1.4本文研究方法与技术路线 2
2表达系统的类型与特性分析 2
2.1原核表达系统的优劣势评估 2
2.2真核表达系统的适用性探讨 3
2.3植物表达系统的潜力与挑战 3
2.4不同表达系统的比较与选择依据 4
2.5特定重组蛋白的表达系统匹配 4
3影响表达效率的关键因素分析 5
3.1宿主细胞的选择与改造策略 5
3.2诱导条件对表达效率的影响 5
3.3融合标签的设计与功能优化 6
3.4发酵工艺参数的调控机制 6
3.5蛋白折叠与分泌途径的优化 7
4优化策略的具体实施与案例分析 7
4.1高通量筛选技术的应用实践 7
4.2合成生物学在表达系统中的应用 8
4.3工程化宿主菌株的构建实例 8
4.4重组蛋白纯化工艺的优化方案 9
4.5成功案例的经验总结与启示 9
结论 10
参考文献 11
致 谢 12
