Direct labelling of proteins using Affimer-conjugate warheads for imaging

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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.