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Single molecule protein sequencing using biological nanopores

Project description

A novel method for protein sequencing

Post-translational modifications (PTM) endow proteins with extensive functional variation paramount for living organisms. However, detection of these modifications by sequencing or mass spectrometry comes with certain limitations. Both techniques are labour-intensive, require extensive sample preparation and cannot account for low-abundance proteins or provide information at the single-cell level. Scientists of the EU-funded SMPSBN project propose to develop a method similar to nanopore DNA sequencing for determining PTMs in a high-throughput manner and with single-molecule sensitivity. Once materialised, this technology is expected to revolutionise proteomics research and pave the way for its use in the clinic.

Objective

Despite the importance of post-translational variation to the function of proteins in a living organism, sequencing proteins to study these variations is still a costly and time-consuming process, with intrinsic limitations. Protein sequencing and detection of post-translational modifications (PTMs) by mass spectrometry, the current gold standard, requires extensive sample preparation and large sample sizes, and possesses a limited dynamic range with respect to sample concentration. These limitations severely restrict its application to biological and clinical problems. A robust method for sequencing proteins and detecting PTMs at the single-molecule level would be revolutionary for proteomics research, allowing biologists to quantify low-abundance proteins as well as distributions and correlations of PTM patterns, all at a single-cell level. I propose a first-of-kind method for protein sequencing with applications in fundamental biology, cancer immunotherapy, and pharmaceutical development. This method uses biological nanopores in a manner similar to nanopore DNA sequencing, an established single-molecule sequencing technology capable of high throughput and single-molecule sensitivity. Developing this method for protein sequencing comes with many significant challenges, which will be addressed over the course of the proposed research.

Coordinator

TECHNISCHE UNIVERSITEIT DELFT
Net EU contribution
€ 187 572,48
Address
STEVINWEG 1
2628 CN Delft
Netherlands

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Region
West-Nederland Zuid-Holland Delft en Westland
Activity type
Higher or Secondary Education Establishments
Links
Total cost
€ 187 572,48