Action in solution: Embedding new technology and new capability in Biomolecular Interactions in the University of Leeds

Life's processes depend on the interactions of macromolecules with ligands and other macromolecules, and the resulting conformational and functional changes that occur upon complex formation. While measuring these interactions has yielded key mechanistic insights over recent decades, including parameters such as binding affinity and cooperativity, detailed measurements typically require well-behaved (monodisperse) samples, significant quantities of material, and simple buffer systems, rather than the complex milieu of physiological environments.

Studies of intrinsically-disordered proteins (IDPs) and self-assembling systems are particularly challenging, but are involved in fascinating and biologically important mechanisms such as viral assembly, liquid-liquid phase separation (LLPS), protein aggregation and amyloid formation.

Here we propose to purchase a Fida-1 instrument which has revolutionised measurements of binding, conformational change and self-assembly of even the most complex systems. Fida-1, supplied by Fidabio, offers real-time, sensitive detection of molecules coupled with rapid microfluidic separation (Flow-Induced Dispersion Analysis) (FIDA). Fida-1 is unique in its capability to resolve size, conformation and binding effects in polydisperse samples, bringing clarity to complex and self-assembling systems that have confounded existing techniques (e.g. dynamic or static light scattering).

Fida-1 can resolve mixtures with a broad range of hydrodynamic radii, in ensemble mode for smaller particles and via simultaneous single-particle detection of larger assemblies. Experiments are performed in solution, and can include label-free measurements of virtually any system. It is automated, takes vials or 96-well plates, and has a uniquely broad size range, including small molecules, nucleic acids, proteins/peptides, polysaccharides, fibrils, viruses, biomolecular condensates and other nanoparticles, including membrane vesicles and virus-like particles.

Low concentrations can be measured, in minutes, using mL of material, in physiological milieu and without need for purification.

Fida-1 will be widely utilised in projects spanning the biosciences and biotechnology, from plants and microbes to man, from biophysics to cell biology, from health to disease. Specifically, we will use the instrument for sample quality control, enhancing efficiency of high-resolution structural analysis, binding-induced conformational changes, especially for dynamic biomolecules in complex environments.

Fida-1 will be the first instrument of its type in a UK University, available to users across the Leeds campus, as well as academia and industry from the UK and beyond. Our AIM is to transform our ability to monitor and analyse macromolecular complexes key to life's processes and enhance capability in allied, high-resolution structural methods at the heart of the Astbury Centre for Structural Molecular Biology (ACSMB).

Focusing on projects including LLPS, virus assembly, amyloid formation, molecular chaperones, IDPs and polysaccharides, our OBJECTIVE is to bring new capability that will increase fundamental understanding of biological mechanism and its translation. Working closely with the instrument manufacturers, we will run training events and host an annual symposium for Fida-1 users, providing the UK with a hub for exploitation and development of FIDA into the future.