Exceptional yield recombinant vesicle packaged protein export system for research and industrial biotechnology applications.

细胞使用小的膜结合的信封，称为囊泡，来存储和输出细胞外的分子。对细胞进行重新编程以控制这一过程的能力对合成生物学具有巨大的潜力，因为它将使我们能够利用细胞的机械来控制包装和释放被整齐包装成囊泡的具有商业价值的分子。这个项目是基于我们发现了一种劫持细胞的方法来控制这一过程。这项简单而经济的发明是通过在蛋白质上添加一个小标签来实现的。这种标记导致细菌产生大量这种充满了标记的感兴趣分子的膜包装。这些包装从细胞输出到生长发酵液中，导致蛋白质产量增加100倍以上。蛋白质功能在这些包装中保存了几个月。这项新技术代表着重组蛋白生产的一项重大突破，因为它促进了用于生物技术和医疗应用的各种蛋白质的简单、高效和快速纯化。在这个项目中，我们将开发这项技术，并将其应用于提高一系列生物技术和医学上重要蛋白质的生产和纯化。与业界(Fujifilm-DiSynth BioTechnologies)合作，我们将优化系统，以实现大规模发酵培养，并开发方法，使泡囊包装蛋白的纯化变得简单且经济高效。最后，我们将把这种重组囊泡包装系统扩展到更复杂类型的细胞，以进一步扩大治疗蛋白的范围和下游应用。这项技术的发展将有利于发现科学和商业应用，因为它将使科学家能够生产生物分子，如诊断和治疗抗体，由细胞在稳定的环境中输出，以便更容易地分离和储存蛋白质。

Cells use small membrane bound envelopes, called vesicles, to store and export molecules out of the cell. The ability to reprogram a cell to control this process has huge potential for synthetic biology as it would allow us to harness the cell's machinery for the controlled packaging and release of commercially-valuable molecules neatly packaged into vesicles. This project is based on our discovery of a way to hijack the cell to control this process. This simple and cost-effective invention works by adding a small tag onto the protein. This tag results in bacteria producing huge numbers of these membrane packages filled with the tagged molecules of interest. These packages are exported from the cell into the growth broth, and result in more than a 100-fold increase in protein yield. Protein function is preserved within these packages for months in the fridge. This novel technology represents a major breakthrough in recombinant protein production as it facilitates simple, efficient and rapid purification of diverse proteins for use in biotechnology and medical applications. In this project we will develop this technology and apply it to enhance production and purification of a wide range of biotechnologically and medically important proteins. Working with industry (Fujifilm-Diosynth Biotechnologies) we will optimise the system to large scale fermentation cultures, and develop methods to allow simple and cost-effective purification of the vesicle packaged proteins. Finally, we will extend this recombinant vesicle packaging system to more complex types of cell to further extend the range of therapeutic proteins and downstream applications. Development of this technology will benefit both discovery science and commercial applications, as it will allow scientists to produce biomolecules such as diagnostic and therapeutic antibodies, exported by the cells in a stable environment for easier isolation and storage of proteins.