Direct labelling of proteins using Affimer-conjugate warheads for imaging

利用显微镜技术放大肉眼看不见的物体的能力给科学带来了革命性的变化。显微镜已被广泛用于研究生物体，并提高了了解健康和疾病过程的能力。利用抗体检测生物体中单个物体的能力进一步突破了界限，使研究更小的物体（如蛋白质）成为可能。蛋白质调节着身体的每一个过程，有超过20000种不同的蛋白质。抗体通常用于每个分子和细胞生物学实验室，以间接地可视化蛋白质，但随着显微镜技术的改进和“超分辨率”，抗体的大小已成为限制因素之一。为了克服这个问题，超分辨率显微镜采用了更小的探针，通常被称为非抗体结合蛋白，使检测试剂更接近蛋白质。然而，这仍然是一种间接的蛋白质可视化方法，随着超分辨率技术的改进，这些将成为提高分辨率的下一个限制因素。该项目旨在开发一种直接标记目标蛋白的可视化方法。这将利用最近开发的一种称为Affimers的非抗体结合蛋白来传递检测试剂，并将其化学偶联到目标蛋白上。这将提供一个试剂的第一个例子，直接结合成像试剂到内源性蛋白质的可视化和超分辨率显微镜，代表了一个令人兴奋的方法，可以适用于许多其他技术。

The ability to magnify objects that are invisible to the naked eye using a technique called microscopy revolutionised science. Microscopy has been widely used to study organisms and has increased the ability to understand processes in health and disease. The ability to detect individual objects in organisms using antibodies has further pushed the bounderies and enabled the study of even smaller objects, such as proteins. Proteins regulate every process in the body, and there are over 20,000 different proteins. Antibodies are routinely used in every molecular and cellular biology laboratory to indirectly visualise proteins but as microscopy technologies have improved and become 'super-resolution' the size of the antibody has become one of the limiting factors. To overcome this, super-resolution microscopy has adopted using smaller probes, often referred to an non-antibody binding proteins, that places the detection reagent closer to the protein. However, this is still an indirect method of visualising proteins and as super-resolution techniques improve these will become the next limiting factor to increasing resolution. This project aims to develop a method of directly labelling the target protein for visualisation. This will utilise a recently developed non-antibody binding proteins, called Affimers, to deliver the detection reagent and chemically conjugate it to the target protein. This will provide the first example of a reagent to directly conjugate imaging reagents to endogenous proteins for visuallisation and super-resolution microscopy and represents and exciting method that could be adapted for many other techniques.