Optimisation of sortase-mediated protein labelling as a tool for biotechnology and pharmaceutical development
Lead Research Organisation:
University of Leeds
Department Name: Sch of Chemistry
Abstract
Labelling of proteins is fundamental to laboratory investigations and industrial applications such as the development of diagnostics. The market for protein labelling reagents is worth approximately $2bn a year and new innovations which increase the efficiency of protein labelling and reduce the cost of this process are always needed. In this project, we are developing a new protocol for the use of sortase in protein labelling. This technology has the potential to allow the rapid (<1 hour) generation of homogenously modified proteins with minimal excess reagent and small quantities of the labelling protein. During the project we will investigate a range of potential commercial applications including the potential of the technology for application in the generation of modified antibodies, which represent a major new class of anti-cancer drugs.
Publications
Arnott Z
(2024)
Quantitative N- or C-Terminal Labelling of Proteins with Unactivated Peptides by Use of Sortases and a d -Aminopeptidase
in Angewandte Chemie
Arnott ZLP
(2024)
Quantitative N- or C-Terminal Labelling of Proteins with Unactivated Peptides by Use of Sortases and a d-Aminopeptidase.
in Angewandte Chemie (International ed. in English)
Dolan JP
(2022)
Synthesis of cholera toxin B subunit glycoconjugates using site-specific orthogonal oxime and sortase ligation reactions.
in Frontiers in chemistry
Machin DC
(2024)
Sortase-Modified Cholera Toxoids Show Specific Golgi Localization.
in Toxins
Morgan HE
(2022)
Challenges in the use of sortase and other peptide ligases for site-specific protein modification.
in Chemical Society reviews
Morgan HE
(2022)
Combined Application of Orthogonal Sortases and Depsipeptide Substrates for Dual Protein Labeling.
in Bioconjugate chemistry
| Description | In this proposal we continued the development of a protein labelling technology to proof-of-concept stage. Through this project we demonstrated the full scope of the technology and obtained key data that is vital for intellectual property purposes. We have now shown successful high-yielding reaction on several different protein systems and have identified key parameters required for optimisation as well as identifying the key conditions required for successful labelling of proteins. We are now moving to the next phase of the project to identify mechanisms to exploit the results of the project most effectively by collaboration with a targeted industrial partner using internal UoL funding to facilitate this interaciton. This evaluation is still ongoing having been delayed due to the COVID-19 pandemic. |
| Exploitation Route | Prior to publication, we are investigating patent protection of the technology. After this point we anticipate that the technology could be adapted for application in industry or in an academic setting. |
| Sectors | Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
| Description | The technology developed through this proposal is now being used in industrial collaborations with existing and prospective non-academic research partners. In the last year, we have participated in the iCure Discover and iCure Explore schemes to determine market interest and have filed a patent covering the technology developed through this grant. This patent has not yet been published. |
| First Year Of Impact | 2019 |
| Sector | Chemicals,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Economic |
| Description | ACROPATH: Artificial Cells for highly sensitive and RObust diagnosis of PATHogen infections |
| Amount | £1,242,474 (GBP) |
| Funding ID | UKRI/MR/B000291/1 |
| Organisation | Medical Research Council (MRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 02/2025 |
| End | 02/2028 |
| Description | IMProGlyco |
| Amount | € 2,998,000 (EUR) |
| Funding ID | 101186774 |
| Organisation | European Innovation Council |
| Sector | Public |
| Country | European Union (EU) |
| Start | 02/2025 |
| End | 01/2028 |
| Description | Multivalent display of sugar antigens via site selective enzymatic modification of cholera toxin |
| Amount | £112,000 (GBP) |
| Organisation | GlaxoSmithKline (GSK) |
| Sector | Private |
| Country | Global |
| Start | 09/2020 |
| End | 09/2024 |
| Description | Sortase conjugate vaccines |
| Amount | £39,288 (GBP) |
| Organisation | University of Leeds |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 09/2019 |
| End | 03/2020 |
| Title | Method of Trasnpeptidastion |
| Description | A method for the efficient labelling of proteins using transpeptidases. |
| IP Reference | |
| Protection | Patent / Patent application |
| Year Protection Granted | 2023 |
| Licensed | No |