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Nanopore spectrometry for the detection of proteins and their modifications

PhD ceremony:R.C.A. Versloot, MSc
When:March 07, 2023
Start:14:30
Supervisor:G. (Giovanni) Maglia, Prof
Co-supervisor:dr. C. Wloka
Where:Academy building RUG
Faculty:Science and Engineering
Nanopore spectrometry for the detection of proteins and their
modifications

Our understanding of the cell, the most basic functional unit of life, has evolved rapidly since the sequencing of the human genome in 2000. DNA sequencing is now a well-established technique and has greatly increased our understanding of the blueprint of life. DNA alone, however, cannot explain the heterogeneity of cells within an organism and the rapid change of protein expression levels due to external stimuli. The proteome, the protein content of cells and tissues, is directly linked to cell functioning and alterations at the proteome level are relevant for prevalent diseases including cancer and Alzheimer’s disease. Information about the proteome could potentially be used for personalised medicine, where the treatment is optimised for a patient’s unique biochemistry. Proteomics, the study of the proteome, is therefore an active field of research, that currently relies on mass spectrometry for the analysis of the protein content in samples. Mass spectrometry is a powerful technique for the detection of proteins in complex samples, but often fails to detect small nuances in the protein content, that could potentially be detected by more accurate, single-molecule techniques. In this thesis, we focus on the adaptability of biological nanopores as powerful single-molecule sensors for the detection and identification of peptides and proteins, as well as their post-translational modifications. We show that nanopores can modified to increase the interaction with peptides. The modified nanopores can detect and identify mixtures of peptides as well as their modifications.