BEACON - A multi-user and multi-project facility for biological macromolecules and nanoparticles characterization in solution

Biological entities are composed of small molecules (e.g., water), macromolecules (proteins, nucleic acids, lipids, polysaccharides), supramolecular assemblies or macromolecular complexes, which assemble into subcellular organelles and cells, which in turn form supracellular systems such as tissues and organs. One of the key features of biological systems is the complex, specific or non-specific, intra- and inter-molecular interactions, which determine the structure and, further, the biological functions associated with the structure. As an example, hemoglobin is a tetrameric protein that binds and transports four oxygen molecules per unit and then releases them to myoglobin. The contacts between biological molecules, or biomolecular interactions, are also responsible for pathogen invasion and various diseases. SARS-CoV-2 virus binds to ACE2, the host target cell receptor, through the receptor binding domain of its spike protein. Neurodegenerative diseases such as Alzheimer's or Parkinson's are associated with the prion-like propagation and aggregation of toxic proteins. The ability to measure key physical parameters (e.g., mass, size, shape, interaction, and conjugation) of biological macromolecules and nanoparticles in solution will answer many FUNDAMENTAL QUESTIONS IN BIOLOGY and help us to understand THE RULES OF LIFE. Not only will the answers and the rules help to advance biotechnology and biological science, they guide our solutions to the grand challenges facing humanity (e.g., new and re-emerging diseases, epidemic, climate change, and sustainability etc). The aim of this proposal is to purchase a state-of-the-art equipment to enable biophysical measurements. The equipment will support a host of research projects underpinning BBSRC priorities, which neatly align with the visions of the four flagship institutes at the University of Sheffield (Healthy Lifespan, Neuroscience, Energy, and Sustainable Food). On top of that, the equipment allows the University COVID-19 response team to measure the quality of and quantify the biological reagents manufactured at the University to support diagnostic testing of coronavirus. The equipment also enhances our response and preparedness for current and future emergency. This equipment will be maintained by a dedicated team in the Department of Chemical & Biological Engineering at the University.

The principal aim is to acquire an SEC|AF4|CG-MALS-DLS equipment for the University of Sheffield to conduct advanced biophysical characterization of macromolecules and nanoparticles in solution. The equipment combines the separation/mixing technology (SEC, AF4 and CG) and the light scattering technology (MALS and DLS). SEC and AF4 are 2-phase separation and 1-phase separation techniques, respectively. CG is a mixing technology using a multi-syringe pump system. It can be programmed to provide stop-flow injections at a series of well-defined concentrations or compositions. Through module arrangement, the equipment enables its users to perform three types of analyses: SEC-MALS-DLS, AF4-MALS-DLS and CG-MALS-DLS. This state-of-the-art equipment allows us to measure molar mass, size (RMS radius and hydrodynamic radius), conformation (shape, structure and branching parameters), interaction (binding affinity and stoichiometry), and conjugation (molecular weight and fraction of each constituent in a conjugate). The analysis can be performed on proteins, peptides, biopharmaceuticals, biopolymers, and nanoparticles (e.g., micelles, viruses, liposomes, polymersomes, and nanocapsules etc). Information derived from these measurements include native properties (oligomeric state), solution quality, size distribution, aggregation, protein stability, and protein unfolding, to name a few. This SEC|AF4|CG-MALS-DLS equipment will support a host of research projects underpinning BBSRC priorities. These projects include COVID-19 research, biopharmaceutical manufacturing, bioplastic production, RNA nuclear export, plant cell wall, amyloid formation, energy homeostasis, antibacterial drug, b-amino acids, crop protection, membrane remodelling, and high-cell-density cultivation of bacteria. These research activities align with the four flagship research institutes of the University of Sheffield (Healthy Lifespan, Neuroscience, Energy and Sustainable Food).