UKAMPRO Research
We focus on supporting basic and clinical research in the Centre, developing new approaches to optimise amyloid identification and quantitation and collaborations with groups both on and off-site.
Our large clinical proteomics database (>2500 samples) allows an in-depth analysis of the data. Recent findings:
- Formalin fixing can result in partial lysine methylation of wild type transthyretin and the misidentification of variant p.V122I.
- Fibrinogen Aa amyloid can be distinguished from thrombus-derived FibAa using variant searches and the relative scores of the b and g chains.
- ApoA-IV amyloid is associated with presence of fibrillogenic signal sequence.
We are an active member of the European Proteomics Amyloid Network (EPAN), established in London in 2017, aiming to define standard procedures and share experiences on amyloid proteomics and related technologies.
Partners
| Institution | Location |
|---|---|
| Centre Hospitalier Universitaire de Toulouse | Toulouse, France |
| Universitätsklinikum Schleswig-Holstein | Kiel, Germany |
| Università degli Studi di Pavia | Pavia, Italy |
| Fondazione IRCCS Policlinico San Matteo | Pavia, Italy |
| Istituto di Tecnologie Biomediche - CNR | Milan, Italy |
| Rijksuniversiteit Groningen | Groningen, Netherlands |
| Intercollegiate Faculty of Biotechnology UG & MUG | Gdansk, Poland |
| Centro Nacional de Investigaciones Cardiovasculares | Madrid, Spain |
| Uppsala Universitet | Uppsala, Sweden |
| ETH Zürich | Zurich, Switzerland |
| UCL Centre for Amyloidosis | London, UK |
For external and collaborative studies
Including amyloid fibril extraction, 1D gel analysis, non-human samples, protein MW analysis, protein purification, PTM analysis and general advice.
Activities
MS data exchange
In the context of WG1, successful comparison has been obtained from the MS data exchange working group. MS data were shared between London (UCL-NAC) and Milan (ITB-CNR) facilities and the results showed very good reproducibility in terms of amyloid identification. An inter‐centre validation study was carried out comparing proteomics data obtained through different software platforms and bioinformatics tools at London (UCL-NAC) and Milan (ITB-CNR) centres. The work showed a high concordance (>92%) between the proteomics data (Canetti D et al, Molecules. 2021).
Database preparation
In the context of WG3 the activity concerns the reconstruction of a database containing unique immunoglobulin light chain from AL patients. To reach this goal, we are developing a computational tool to automatically compare, classify, update and store the immunoglobulin light chain sequences available from the major public databases, including Uniprot, NCBI and ALbase. The resulting not redundant database will be useful for diagnostic and profiling purposes relying on mass spectrometry-based proteomic approaches.
Transfer of best practices
MALDI imaging of amyloid at Kiel showed that Vitronectin is intimately associated with amyloid deposits (Winter et al, J Histochem Cytochem. 2015). Proteomics data from UCL in London were examined and showed good relationship between the Mascot scores of Vitronectin and the accepted amyloid signatures (ApoA-IV, ApoE and SAP). This data is also in agreement with the work performed from the ITB-CNR group (Brambilla et al, Blood. 2012).
Funding / Acknowledgements
UKAMPRO is a joint venture between the UCL Wolfson Drug Discovery Unit and the NHS National Amyloidosis Centre.
Development was funded through the UK National Institute for Health Research Biomedical Research Centre and Unit Funding scheme via the UCLH/University College London Biomedical Research Centre, the UCL Technology Fund and direct funds from NHS England.
The mass spectrometer was funded through generous grants from the Wolfson Foundation and the UCL Amyloidosis Research Fund.
Clinical samples were collected and analysed as part of routine clinical examination. Informed consent was obtained from all patients and clinical care was in accordance with the Declaration of Helsinki.
