Bio-nanomanufacturing of Protein Therapeutics Using Membrane Microfluidics

使用膜微流体的蛋白质治疗药物的生物纳米制造

• 批准号: 1728049
• 负责人: Susan Daniel
• 金额: $ 37.44万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1728049&HistoricalAwards=false
• 关键词: Bio; nanomanufacturing; Protein; Therapeutics; Using

The market for therapeutic proteins is valued near $140 billion annually. Many of these therapeutics are glycoproteins, which require the addition of specific sugars, called 'glycans' at an exact position on the protein through a process called protein glycosylation. The glycan affects protein folding and function and ensures it retains its therapeutic potency. In nature, glycoproteins are produced through a series of sequential reactions inside a cell. Making therapeutic glycoproteins within cells is challenging for a variety of reasons, and extensive and costly purification steps are required to harvest the therapeutic material. With this award, a cell-free glycosylation network will be constructed in a microfluidic device that separates reactions in space and time, giving supreme flexibility in optimizing individual reactions and constructing glycans with high specificity. The benefits of this manufacturing paradigm to society are reducing the cost of these drugs and providing scientists an avenue to design and develop synthetic drug compounds that may or may not exist in nature to treat disease. The related education plan creates a hands-on bio-nanomanufacturing activity for a high school girls organized by the PIs and their student trainees, so that young women will understand the power of biotechnology and be inspired to pursue these career paths. Cell-free protein synthesis holds great promise for producing high-value, biotherapeutic nanomaterials without cell culture and benefitting from chemical manufacturing know-how. Here, raw materials and biological enzymes are mixed to produce biological products. Shortcomings of this approach are competing reactions, side products, and low yields. Cells avoid these shortcomings by localizing reactions within subcellular compartments and orchestrating the reaction sequences. The biocatalysts that give the final molecule its essential posttranslational features are compartmentalized in membranes. Handling enzymes outside of their native lipid environment can drastically reduce their activity. Thus, in vitro, sequential, bio-enzymatic reactions have never been achieved in a cell-free manner. The research objective is to mimic the elegant compartmentalization strategies used by cells in a microfluidic biomembrane device that organizes biological reactions in proper spatial and temporal sequence. These devices will generate authentically glycosylated proteins. Through assessment of nanostructure product architectures, this work will advance understanding of nanoscale phenomena and processes for nanomaterials manufacture and discovery. This cell-free device concept will enable facile optimization of glycosylated protein production, and provide a framework for understanding how experimental conditions affect product yield and quality that is broadly applicable to the bio-nanomanufacturing of virtually any posttranslationally-modified protein.

治疗性蛋白质的市场价值每年接近1400亿美元。这些疗法中的许多都是糖蛋白，需要通过一种名为蛋白质糖基化的过程在蛋白质上的确切位置添加特定的糖，即所谓的“多糖”。多糖影响蛋白质的折叠和功能，并确保其保持其治疗效力。在自然界中，糖蛋白是通过细胞内的一系列连续反应产生的。由于各种原因，在细胞内制造治疗性糖蛋白是具有挑战性的，需要广泛而昂贵的纯化步骤来获得治疗性材料。获得该奖项后，将在微流控设备中构建无细胞糖基化网络，该设备将在空间和时间上分离反应，在优化个别反应和构建具有高度特异性的多糖方面给予极大的灵活性。这种制造模式给社会带来的好处是降低了这些药物的成本，并为科学家提供了设计和开发合成药物化合物的途径，这些化合物可能存在于自然界中，也可能不存在于治疗疾病的自然界中。相关的教育计划为高中女生组织了一项亲身实践的生物纳米制造活动，由私人投资机构及其学生实习生组织，让年轻女性了解生物技术的力量，并受到激励，追求这些职业道路。无细胞蛋白质合成在无需细胞培养和受益于化学制造技术的情况下生产高价值的生物治疗纳米材料具有很大的前景。在这里，原料和生物酶混合在一起生产生物制品。这种方法的缺点是竞争反应、副产物和低产率。细胞通过在亚细胞间隔内定位反应并协调反应序列来避免这些缺点。生物催化剂使最终的分子具有翻译后的基本特征，这些生物催化剂被分隔在膜中。在天然脂质环境之外处理酶会极大地降低它们的活性。因此，在体外，连续的生物酶反应从来没有以无细胞的方式实现过。该研究的目标是模仿微流控生物膜装置中细胞所使用的优雅的区隔策略，该装置以适当的空间和时间顺序组织生物反应。这些设备将产生真正的糖基化蛋白质。通过对纳米结构产品结构的评估，这项工作将促进对纳米材料制造和发现的纳米级现象和过程的理解。这种无细胞设备的概念将使糖基化蛋白质生产得到便捷的优化，并为理解实验条件如何影响产品产量和质量提供了一个框架，该框架广泛适用于几乎任何翻译后修饰蛋白质的生物纳米制造。