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

Lead Research Organisation: University of Sheffield
Department Name: Chemical & Biological Engineering

Abstract

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.

Technical Summary

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).

Publications

10 25 50
 
Description (1) We have developed an analytical workflow to analyse virus particles using the FluMist samples provided by AstraZeneca.
Exploitation Route AstraZeneca is keen to continue the partnership with Sheffield, and to use biophysical method to optimise biomanufacturing of FluMist.
Sectors Healthcare

Manufacturing

including Industrial Biotechology

Pharmaceuticals and Medical Biotechnology

 
Description We are very closed to spinning out a company called ReNewFood (we are finalizing the paperwork required for incorporation).
Sector Manufacturing, including Industrial Biotechology
Impact Types Societal

Economic

 
Description ReNewFood
Amount £19,800 (GBP)
Funding ID NXW- EX-JAN23-02- SHEFFIELD 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 01/2023 
End 04/2023
 
Description ReNewFood (IPDaC fund)
Amount £52,882 (GBP)
Funding ID X/185198 
Organisation University of Sheffield 
Sector Academic/University
Country United Kingdom
Start 12/2023 
End 09/2024
 
Description ReNewFood (IPDaC fund)
Amount £4,760 (GBP)
Funding ID X/015411 
Organisation University of Sheffield 
Sector Academic/University
Country United Kingdom
Start 02/2023 
End 07/2023
 
Description ReNewFood (IPDaC fund)
Amount £47,837 (GBP)
Funding ID X/015414 
Organisation University of Sheffield 
Sector Academic/University
Country United Kingdom
Start 02/2023 
End 04/2024
 
Description UK-South East Asia Vaccine Manufacturing Research Hub
Amount £7,600,000 (GBP)
Funding ID EP/Y009894/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 11/2023 
End 03/2028
 
Description Vaccine development in partnership with LMIC countries (ISPF ODA funding)
Amount £100,000 (GBP)
Funding ID RE-CL-2023-09 
Organisation United Kingdom Research and Innovation 
Department Research England
Sector Public
Country United Kingdom
Start 01/2024 
End 03/2024
 
Description 2nd collaborative project with AstraZeneca 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution Myself and Dr. Adam Brown wrote the proposal on Synthetic Biology-Based Polymerase and Promoter Engineering for Production of Next-Generation mRNA Therapeutics
Collaborator Contribution (1) BBSRC iCASE studentship (2) £80000 cash contribution (3) Industrial placement for PhD student
Impact PhD student (Alessandra Sabatini) has started her PhD in Oct 2023.
Start Year 2023
 
Description 3rd collaborative project with AstraZeneca 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution I proposed to use BEACON suite to study influenza virus to improve the FluMist manufacturing process.
Collaborator Contribution Provision of influenza virus samples for analysis.
Impact We were able to produce excellent biophysical data using the samples from AstraZeneca. We are now discussing mechanisms to sustain this collaboration.
Start Year 2022
 
Description 4th collaborative project with AstraZeneca 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution We have established analytical workflow to analyze virus and virus-like particles.
Collaborator Contribution AstraZeneca is interested in virus-like particle (VLP) for vaccine development and manufacturing.
Impact AstraZeneca is now considering part-funding a PhD studentship.
Start Year 2024
 
Description Partnership with AZ Cambridge 
Organisation AstraZeneca
Country United Kingdom 
Sector Private 
PI Contribution Myself and Dr. Barbara Ciani (coI in this grant) formulated the research proposal to use BEACON suite to study coformulation of biologics.
Collaborator Contribution (1) AZ will provide cash contribution of £50000 to the project (2) AZ will host the PhD student involved for 3 months and cover the expenses incurred (£30000) (3) AZ will provide biologics for coformulation study (in-kind contribution of £600000)
Impact PhD student (Anyal Settle) is now working on the project.
Start Year 2022