An innovative approach to 'printing' functional protein microarrays from RNA microarrays.

蛋白质是所有活细胞的基本构件，对有机体的正常运作至关重要。了解蛋白质如何相互作用，以及如何与其他生物分子相互作用，是所有生物学研究的核心，对健康和环境领域的科学进步具有明显的影响。例如，新的治疗方法和更有效的生物能源产生都依赖于理解和利用蛋白质相互作用。因此，开发研究蛋白质相互作用的工具是BBSRC战略计划中的关键优先事项也就不足为奇了。研究蛋白质相互作用最有效的方法之一是生成一个包含数百到数千个蛋白质的单一表面，这些蛋白质都可以在一个步骤中进行相互作用测试。这种类型的表面被称为功能蛋白质微阵列，可用于进行高通量相互作用研究。考虑到功能蛋白质微阵列在现实世界中的应用，仅在医疗领域，它们就有可能通过在药物发现、疾病诊断和医学筛查中的使用来支持更好的健康。不幸的是，到目前为止，成功创建功能蛋白质微阵列尤其具有挑战性，因此它们未能产生预期的影响。本申请寻求资金用于试点研究，以证明产生功能蛋白质微阵列的新概念，克服了当前方法的局限性。这一概念涉及使用表面上的蛋白质前体(RNA分子)阵列，称为RNA微阵列，在专门准备的面向表面上产生相应的蛋白质微阵列。实验安排包括将两个表面以三明治的形式相互相对放置，中间有一种特定的生物溶液，将RNA分子转化为蛋白质分子。利用一种新的化学步骤，新形成的蛋白质分子在溶液中附着到专门准备的面向表面，形成功能蛋白质微阵列。为了证明这一新概念，该项目将依次演示这一过程的每个阶段，然后将其结合在一起，通过有效地从前体RNA微阵列中打印出来，创建一个功能蛋白质微阵列。这项工作是创新的、及时的和多学科的，使用了化学上的最新进展，以促进蛋白质附着到微阵列载玻片表面，以及我们最近最先进的产生前体RNA微阵列的专利技术。重要的是，这项应用不是关于这种RNA微阵列技术的渐进式进一步发展，而是关于利用它并证明在一个完全独立的领域中的新工具的新概念；特别是用于产生功能性蛋白质微阵列。这项研究的潜力很大，通过以简单有效的方式创建具有更强稳健性、更小的斑点大小和不受限制的蛋白质大小的功能蛋白质微阵列，提供了能力上的阶梯变化。这克服了现有功能蛋白质微阵列技术的关键限制，并释放了最初预测的巨大好处。

Proteins are the fundamental building blocks of all living cells and are essential for the proper functioning of an organism. Understanding how proteins interact with each other, and with other biological molecules, lies at the heart of all biological research and has clear implications for scientific progress within both health and environmental fields. For example, new therapeutics and more efficient bioenergy generation both rely on understanding and exploiting protein interactions. It is therefore unsurprising that developing tools to study protein interactions is a key priority within the BBSRC strategic plan.One of the most efficient ways of investigating protein interactions is to generate a single surface containing hundreds-to-thousands of proteins, which can all be tested for interactions in one step. A surface of this type is known as a functional protein microarray and can be used to conduct high throughput interaction studies. Considering the real world applications of functional protein microarrays, within the medical arena alone, they have the potential to underpin better health through their use in drug discovery, disease diagnosis and medical screening. Unfortunately, to date, the successful creation of functional protein microarrays has been particularly challenging and they have therefore failed to deliver the impact anticipated.This application seeks funding for pilot research to demonstrate a novel concept for generating functional protein microarrays that overcomes the limitations of the current approaches. The concept involves using an array of protein precursors (RNA molecules) on a surface, known as an RNA microarray, to generate a corresponding protein microarray on a specially prepared facing surface. The experimental setup involves placing the two surfaces opposite each other in a sandwich arrangement, with a specific biological solution in-between that converts the RNA molecules into protein molecules. Using a novel chemistry step, the newly formed protein molecules in solution attach themselves to the specially prepared facing surface, forming the functional protein microarray. To prove the novel concept, this project will involve demonstrating each stage of the process in turn, before bringing it all together to create a functional protein microarray by effectively 'printing' it from the precursor RNA microarray. This work is innovative, timely and multi-disciplinary, employing the latest advances in chemistry, to facilitate the attachment of the proteins to the microarray slide surface, as well as our recent, state-of-the-art, patented technology for generating the precursor RNA microarray. Importantly, this application is not about incremental further development of this RNA microarray technology, but is instead about exploiting it and proving a novel concept for a new tool in a completely separate field; specifically, for the generation of functional protein microarrays. The potential of this research is considerable, offering a step change in capability by creating functional protein microarrays with greater robustness, smaller spot sizes and unrestricted protein sizes, in a simple and efficient manner. This overcomes the key limitations of existing functional protein microarray technologies and unlocks the vast benefits originally forecast.