Protein Vesicles: Understanding Self-Assembly of Fusion Proteins into Vesicles to Engineer Structures and Biofunctional Properties

非技术：该奖项支持由功能性蛋白质制成的囊泡的研究和开发。囊泡是一种微型或纳米级的中空容器，可以装载货物，用于药物输送、生物成像或作为微型生物反应器。功能蛋白可以治疗或执行反应，由这些蛋白制成的囊泡获得其功能。所提出的研究旨在了解蛋白质的性质和制造条件如何影响形成的囊泡的大小和结构，以便人们可以在每种应用中控制这些性质。为了证明它们的用途，将由乳腺癌结合蛋白形成囊泡，并装载化疗药物，并测试它们杀死乳腺癌细胞的能力。总的来说，这项研究将有助于蛋白质囊泡的发展，用于各种支持人类健康的应用，研究结果将发表以供公众传播。在这项工作中，博士后和本科生将接受材料工程和生物学相结合的最先进方法的培训，为他们在工程、生物材料或药物输送领域的职业生涯做好准备。此外，该奖项将支持从中学到博士后教育水平的STEM女性招聘、指导和留住女性的活动，包括在实验室为中学女生进行动手实验。技术：该奖项支持使用光散射和电子显微镜等技术研究融合蛋白自组装成囊泡。所产生的知识将应用于微调囊泡特性，包括大小和膜结构，通过简单的改变组装条件。拟议的研究预计将产生范围从~100纳米到2微米的单层或双层膜的囊泡，同时保持形成囊泡膜的蛋白质的功能和装载货物的能力。功能应用将通过将含有单链抗体片段的融合蛋白组装成囊泡来证明，这些融合蛋白与乳腺癌细胞上的Her2受体结合并携带化疗药物。这些功能性囊泡特异性靶向并杀死Her2+乳腺癌细胞的能力将在体外进行评估。调查结果将公布，供公众传播。在这项工作中，一名博士后和本科生将接受重组蛋白设计和生产、材料表征和细胞培养分析等方法的培训。他们将接受工程、生物材料或药物输送方面的指导和准备。此外，该奖项将支持从中学到博士后的教育水平招募和留住STEM女性的活动，包括在实验室为中学女生进行动手实验。

Non-technical:This award supports the study and development of vesicles made from functional proteins. Vesicles are micro or nano-scale hollow containers that can be loaded with cargo and used for drug delivery, bioimaging, or as tiny bioreactors. Functional proteins can be therapeutic or perform reactions, and vesicles made from these proteins gain their function. The proposed studies are expected in developing an understanding on how the properties of the proteins and the fabrication conditions affect the size and structure of the formed vesicles so that one may control these properties for each application. To demonstrate their use, vesicles will be formed from a breast cancer binding protein and loaded with chemotherapeutic cargo and tested for their ability to kill breast cancer cells. Overall, this research will contribute to the development of protein vesicles for a variety of applications that support human health and the results will be published for public dissemination. During this work, a postdoctoral student and undergraduates will be trained in state of the art methods that integrate materials engineering and biology and they will be prepared for careers in engineering, biomaterials, or drug delivery. Additionally, this award will support activities to recruit, mentor and retain women in STEM at education levels ranging from middle school to post-doctoral, including hands-on experiments for middle school girls in the lab.Technical:This award supports the study of fusion protein self-assembly into vesicles using techniques such as light scattering and electron microscopy. The knowledge generated will be applied to fine tune vesicle properties, including size and membrane structure, through simple changes in assembly conditions. The proposed studies are expected to produce vesicles ranging from ~100 nanometer to 2 micro meter with single or double layer membranes while maintaining the functionality of proteins that form the vesicle membrane and cargo loading ability. Functional application will be demonstrated by assembling vesicles from fusion proteins containing single chain antibody fragments that bind the Her2 receptor on breast cancer cells and carry chemotherapeutic. The ability of these functional vesicles to specifically target and kill Her2+ breast cancer cells will be assessed in vitro. The results will be published for public dissemination. During this work, a postdoctoral student and undergraduates will be trained in methods including recombinant protein design and production, materials characterization, and cell culture assays. They will be mentored and prepared for careers in engineering, biomaterials, or drug delivery. Additionally, this award will support activities to recruit and retain women in STEM at education levels ranging from middle school to post-doctoral, including hands-on experiments for middle school girls in the lab.