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

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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.