Collaborative Research: Advanced biomanufacturing of functional bionanoparticles for biomedical engineering applications

1604826/1604925：雷/陈：新纳米材料的研究在过去十年中经历了爆炸性的增长。然而，从实验室规模向大规模生产过渡的主要挑战，如高通量制造工艺、一致性和大批量产品质量监控方法一直是发挥其巨大潜力的瓶颈。该方案的目标是设计一种先进的制造工艺来制造基因工程多功能生物纳米颗粒(BIO-NPs)，并检验和验证其在脑肿瘤非侵入性成像中的应用。如果成功，这将提供一个极好的示范，从NSF式的基础科学到现实世界的应用。该项目通过将先进的生物制造和生物成像模块整合到他们的教育和实验室培训中，将对研究生、本科生和高中生的教育产生积极的影响。将向学生提供一门新的以研究为导向的生物制造课程。这一多学科项目旨在通过基因工程的纳米囊泡形成的大肠杆菌的发酵合成新型纳米多功能外膜囊泡(OMV)，并将其应用于脑肿瘤的非侵入性生物成像。为了实现这一目标，首先将利用重组DNA技术设计新型的基因工程蛋白多功能生物纳米颗粒，用于捕获和检测生物成像功能。生物纳米粒是基于脂类的OMV，大小均匀，双层的外层小叶上装饰着新型的工程蛋白融合，赋予了多功能。OMV共展示多个拷贝的超活性NanoLuc荧光素酶(比传统的萤火虫或Renilla荧光素酶的活性高约150倍)，将包含(I)用于锚定感兴趣抗体的抗体结合结构域，以及(Ii)用于通过大小过滤简单纯化OMV的热响应弹性蛋白样蛋白结构域。然后，将设计一种集成了两级粒度过滤的发酵工艺来生产多功能OMV。最后，该项目将验证这些OMV用于高性能脑肿瘤生物成像的功能。这项拟议的研究将为生物制造提供一个新的视角，同时该产品可以极大地促进全球公共健康。这种新型的可扩展的基因工程制造平台可以推广到制备具有许多其他所需功能的OMV，适合于广泛的应用，包括生物修复、生物催化剂、生物传感、生物质转化、疫苗和药物输送。

1604826/1604925:Lei/ChenResearch on new nanomaterials has undergone explosive growth in the past decade. However, the main challenges of the transition from laboratory-scale to mass production, such as high throughput manufacturing processes, uniformity, and methodology of monitoring the quality of large-quantity products have been the bottlenecks to realize their tremendous potential. The goal of this proposal is to design an advanced manufacturing process to manufacture genetically engineered multi-functional bio-nanoparticles (bio-NPs) and to examine and validate their utility for non-invasive imaging of brain tumor cancer. If successful, this will provide an excellent demonstration from NSF-style basic science to real-world applications. This project will positively impact education of graduate, undergraduate and high school students by integrating advanced biomanufacturing and bioimaging modules into their educational and laboratory training. A new research-oriented course in Biomanufacturing will be offered to students. This multidisciplinary project aims to synthesize novel nano-sized multi-functional outer membrane vesicles (OMVs)decorated with engineered proteins through fermentation of genetically engineered nano-vesicle-forming E. coli and then apply the decorated OMVs for non-invasive bioimaging of brain tumor. To accomplish this, recombinant DNA technology will first be used to design novel genetically engineered protein multi-functional bio-NPs for capture and detection functions for bioimaging. The bio-NPs are lipid-based OMVs with a uniform size and the outer leaflet of the bilayer is decorated with novel engineered protein fusion, endowing multi-functionality. The OMVs, co-displaying multiple copies of super-active NanoLuc luciferase enzyme (~150-fold more active than that of conventional firefly or Renilla luciferase), will contain (i) an antibody-binding domain for anchoring antibodies of interest, and (ii) a thermo-responsive elastin-like protein domain for simple purification of the OMVs via size filtration. A fermentation process integrated with two-stage size filtration will then be designed for production of multi-functional OMVs. Finally, the project will validate the functionality of these OMVs for high performance bioimaging of brain tumor. The proposed research will offer a new perspective to biomanufacturing while the product can greatly promote global public health. This novel scalable genetically-engineered manufacturing platform can be generalized to prepare the OMVs with many other desired functions suitable for a wide range of applications including bioremediation, biocatalysts, biosensing, biomass conversion, vaccines, and drug delivery.