Unleashing the next-generation single-cell proteomics workflows  

Abstract:

Single-cell proteomics by Mass Spectrometry (scp-MS) can provide valuable insight into distinct cell-states and signalling patterns present in a cell population. However, carrying out proteomics profiling from the limited amount of material encapsulated in an individual cell presents significant challenges. Tremendous efforts have been made to optimize all aspects of scp-MS, with the aim of minimizing losses during sample preparation and maximizing sensitivity of data acquisition. Nevertheless, continuous development is needed to increase both the quantifiable proteome depth per cell, the quantitative accuracy and precision of those measurements, and the number of cells measured per unit time. 

Here, we will present recent approaches developed in Erwin Schoof’s Cell Diversity Lab. We will cover key aspects of the entire workflow, ranging from sample preparation and chromatographic separation, to advanced data acquisition and data interpretation and visualization methods. We will cover a range of instrumentation, including the newly announced ThermoFisher Orbitrap Astral MS, and span both TMTPro-labeled and label-free approaches. By showcasing the application of our methods to biological questions, we aim to convey possible biomedical implications of scp-MS, and provide an overview of the current technological state of the field.  

Biography:

Erwin is a recently appointed Associate Professor in single-cell proteomics in disease biology at the Technical University of Denmark. With a background in Medicinal Chemistry (MSc. from Utrecht University) and molecular systems biology in cancer (PhD from the Technical University of Denmark), his research focuses on normal and malignant hematopoiesis. He dedicated his postdoc to the investigation of healthy and malignant blood stem cells in the lab of John Dick (Toronto, Canada) and Bo Porse (Copenhagen, Denmark), and has spent the last few years optimizing single-cell proteomics workflows to be used for deciphering cellular heterogeneity. He currently heads the Cell Diversity Lab, focusing on the development and application of high-sensitivity proteomics workflows.