Project description
Technological innovations for enhanced protein identification
Characterisation of the proteins produced by a cell or organism can provide fundamental insights into cellular processes and disease mechanisms. Therefore, there is a need for single-molecule protein sensing and sequencing techniques with great sensitivity and specificity. With the support of the Marie Skłodowska-Curie Actions programme, the Plas-ID project aims to overcome existing technical challenges associated with full-length reads and identification of low-abundance proteins through a molecule sensing device that utilises dual-colour amino acid-specific labelling. The device will be designed to slow protein movement and separate them by molecular mass during electrophoresis. The idea is to provide a diagnostic tool for identifying and exploring the role of clinically relevant isoforms of vascular endothelial growth factor.
Objective
Single-molecule protein sensing and sequencing technologies are emerging on the horizon as new powerful tools, which ultimate goal is the characterization of the proteome, namely the total set of proteins made by a cell or organism. Despite recent advances, challenges in terms of full-length reads, high throughput and intrinsic complexity related to the larger set of building blocks (i.e. 20 amino acids) still remain. Furthermore, the unambiguous identification of proteins at low concentration is an open issue for conventional techniques, such as Mass Spectroscopy and ELISA. I aim to develop a plasmon-enhanced single-molecule sensing device to track and identify proteins, combining a dual-color amino-acid-specific technique to label the proteins and the enhancement to the fluorescence provided by properly-designed plasmonic nanostructures integrated within a nanochannel-based device. The plasmonic nanochannel is filled with a custom-made gel to slow down the motion of proteins and separate them by their molecular mass during the electrophoretic migration, meanwhile plasmonic nanostructures enhance the fluorescence signals acquired from each individual dual-color labelled protein. I will focus on the optimization of the device and testing its capabilities for the discrimination of clinically-relevant isoforms of Vascular endothelial growth factor (VEGFa) family. I will tackle the challenge of isoforms identification even further, by using the device to discriminate between VEGFa and VEGFb, which differ for only six amino acids at the C termini. Results from this work will provide a technique for the identification and quantification of protein isoforms with single-molecule resolution. Potentially, the use of this diagnostic tool will provide new insight into the role of VEGF isoforms and their up/down regulation into several diseases and in turns, also new avenues for targeted therapeutics.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomics
- social sciencessociologysocial issuessocial inequalities
- natural sciencesphysical sciencesopticsspectroscopy
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Keywords
Programme(s)
- HORIZON.1.2 - Marie Skłodowska-Curie Actions (MSCA) Main Programme
Funding Scheme
HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European FellowshipsCoordinator
32000 Haifa
Israel