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.
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.
People |
ORCID iD |
Sheena Radford (Primary Supervisor) |
Publications
Cawood EE
(2020)
Modulation of Amyloidogenic Protein Self-Assembly Using Tethered Small Molecules.
in Journal of the American Chemical Society
Ebo JS
(2020)
An in vivo platform to select and evolve aggregation-resistant proteins.
in Nature communications
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 |
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 |