As the field of single-cell proteomics (SCP) advances, new challenges and opportunities emerge in the quest to understand cellular heterogeneity within complex biological systems. One of the primary limitations in SCP is the reproducibility and scalability of sample processing workflows. Recent technological advances in single-cell isolation, such as the CellenONE cell sorter (Cellenion), have significantly improved the efficiency and reproducibility of single-cell collection, enabling more reliable downstream molecular analysis. Despite these advancements, challenges persist, particularly in data acquisition, where instrumentation often requires a delicate balance between sample throughput and the depth of proteome coverage.
At the Monash Proteomics & Metabolomics Platform, we are committed to advancing the capabilities of SCP by optimising experimental workflows including sample preparation techniques, acquisition methods, and improved data analysis pipelines for both label-free and multiplexed approaches. These efforts aim to push the boundaries of SCP in terms of reproducibility, scalability, and proteome depth.
We will present recent developments that highlight our progress in enhancing SCP capabilities using a Thermo Scientific Orbitrap Exploris 480 mass spectrometer. Our work is focused on improving the reproducibility and scalability of workflows while maximising proteome coverage. Ultimately, these advancements are crucial for gaining a deeper understanding of cellular heterogeneity and its implications for complex biological systems. By addressing these challenges, we hope to contribute significantly to the broader field of single-cell proteomics.