Evolving and improving protein stability for enhanced biopharmaceuticals

Lead Research Organisation: University of Leeds
Department Name: Astbury Centre

Abstract

The importance of antibody-based therapeutics is now well established and this expanding sector, in which the UK is a large stakeholder, now has yearly sales of greater than $75 billion. However, the production and formulation of biopharmaceuticals can be problematic, jeopardising their successful development. Specifically, protein self-association results in major hurdles which must be overcome for the translation of a promising candidate to a blockbuster bio-therapeutic and can result in the failure of promising candidate biologics during pre-clinical development. The ability to predict and/or detect sequences prone to aggregation, thus, is the holy grail of the biologics industry.
In this studentship we will develop new understandings of how and why proteins aggregate and which are likely to be unsuitable for bioprocessing and development. We will achieve this using protein engineering to evolve new protein sequences with reduced/enhanced stability and/or aggregation propensity using a split-beta-lactamase system we have recently developed (Foit et al., Molecular Cell, 2009 and Saunders, Brockwell, Radford et al., Nature Chemical Biology, 2016). We will then use sophisticated biophysical methods (e.g. SEC-MALLS, DLS, HX, NMR and FCS) to measure the stability, dynamics and aggregation propensity of the resulting sequences. Finally, we will determine how the bioprocess environment (flow and interaction at interfaces) affects the behaviour of the selected protein sequences (using custom build flow devices developed as part of an on-going collaboration with Professor Nik Kapur (Mechanical Engineering)). We will focus on proteins with an immunoglobulin (Ig) fold, including I27, beta2m (non-aggregating and aggregating model systems) as well as scFvs with known differences in bioprocess behaviour (provided by MedImmune). The model proteins will be studied both as single domains (100 aa), and as polyproteins (mimicking antibody light and heavy chains). These are scaffolds of primary importance in biopharma, including our industrial collaborator. The goal is to discover the mechanisms and determinants of protein self-assembly and why some sequences aggregate with/without flow, whilst others do not.

Publications

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Cawood EE (2020) Modulation of Amyloidogenic Protein Self-Assembly Using Tethered Small Molecules. in Journal of the American Chemical Society

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Ebo JS (2020) Using protein engineering to understand and modulate aggregation. in Current opinion in structural biology

Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M011151/1 30/09/2015 29/09/2023
1774660 Studentship BB/M011151/1 30/09/2016 30/03/2021 Jessica Ebo
 
Description The research has enabled the development of a new method that can screen biopharmaceuticals earlier in their development. The method developed will allow the identification of drugs that are easier to manufacture and can identify the problematic regions of the biopharmaceutical drug buy directed evolution. In the long term this should help companies make drugs faster therefore saving time and money.
Exploitation Route The outcomes will be useful to biopharmaceutical companies wishing to speed up their drug development pipeline to identify the cause of aggregation in biopharmaceuticals. It also has academic interest as the assay can be used to look at proteins involved in disease and further our knowledge on the mechanisms of protein misfolding diseases.
Sectors Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

 
Description ARBRE-MOBIEU network and COST action travel grant for AUC 2017 conference
Amount € 450 (EUR)
Organisation European Cooperation in Science and Technology (COST) 
Sector Public
Country Belgium
Start 06/2017 
End 07/2017
 
Description BioProcess UK conference bursary
Amount £180 (GBP)
Organisation BioIndustry Association 
Sector Charity/Non Profit
Country United Kingdom
Start 11/2018 
End 11/2018
 
Description Biochemical society
Amount £200 (GBP)
Organisation Biochemical Society 
Sector Charity/Non Profit
Country United Kingdom
Start 06/2019 
End 07/2019
 
Description Astbury research retreat research talk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Other audiences
Results and Impact Research presentation at Astbury center research retreat
Year(s) Of Engagement Activity 2019
 
Description PepTalk research presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Speaker at PepTalk protein science week, San Diego, CA, USA
Year(s) Of Engagement Activity 2020
 
Description Protein engineering II conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Research talk at Biochemical society Protein Engineering II conference, York, UK
Year(s) Of Engagement Activity 2019
 
Description White rose DTP postgraduate symposium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact Present research to other postgraduate students and academics in the BBSRC white rose DTP
Year(s) Of Engagement Activity 2019