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。这些治疗剂中的许多是糖蛋白，其需要通过称为蛋白质糖基化的过程在蛋白质上的确切位置添加称为“聚糖”的特定糖。聚糖影响蛋白质折叠和功能，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，在细胞内制备治疗性糖蛋白具有挑战性，并且需要大量且昂贵的纯化步骤来收获治疗性材料。凭借该奖项，将在微流体设备中构建无细胞糖基化网络，该设备在空间和时间上分离反应，在优化单个反应和构建具有高特异性的聚糖方面具有最大的灵活性。这种制造模式对社会的好处是降低了这些药物的成本，并为科学家提供了一种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，也可能不存在于自然界中。相关的教育计划为一名高中女生创建了一个由PI及其学生学员组织的动手生物纳米制造活动，以便年轻女性了解生物技术的力量，并受到激励去追求这些职业道路。无细胞蛋白质合成为生产高价值的生物纳米材料提供了巨大的希望，而无需细胞培养，并受益于化学制造技术。在这里，原料和生物酶混合生产生物制品。这种方法的缺点是竞争反应、副产物和低收率。细胞通过将反应定位在亚细胞区室内并协调反应序列来避免这些缺点。赋予最终分子其基本翻译后特征的生物催化剂被划分在膜中。在其天然脂质环境之外处理酶可以大大降低其活性。因此，体外连续生物酶促反应从未以无细胞方式实现。该研究的目标是模仿细胞在微流体生物膜装置中使用的优雅的区室化策略，该装置以适当的空间和时间顺序组织生物反应。这些装置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现象和纳米材料制造和发现过程的理解。这种无细胞装置概念将使糖基化蛋白质生产的简单优化成为可能，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后修饰蛋白质的生物纳米制造。