"Revolutionizing Proteomics: Introducing the Heat ‘n Beat Method for Rapid, High-Throughput Sample Processing"
This week's Implen NanoPhotometer Journal Club is highlighting the article "Heat ‘n Beat: A Universal High-Throughput End-to-End Proteomics Sample Processing Platform in under an Hour" by Dylan Xavier and colleagues, which presents a significant advancement in the field of proteomics sample preparation. Their work recently published in the journal of analytical chemistry introduced a robust, rapid, and nearly universal protocol for processing small solid tissue samples for mass spectrometry-based proteomic analysis. This new method, referred to as Heat ‘n Beat (HnB), streamlines the preparation process, significantly reducing the time from tissue sample to mass spectrometer loading to under an hour.
Key features of the HnB protocol include a one-tube homogenization and digestion method applicable to various tissue types, irrespective of their preservation methods. The protocol integrates initial washing, tissue disruption, lysis, reduction, alkylation, and proteolysis into a single tube operation, minimizing handling and potential sample loss. This method demonstrates an improved peptide yield, nearly doubling the output compared to previous techniques like the ABLE method, while maintaining high digestion efficiency (85–90%).
HnB's effectiveness was validated across multiple organ types and a large set of human cancer biopsies, showing excellent reproducibility and high-throughput capabilities. For example, 1171 human cancer biopsy samples were processed in 92 hours, yielding consistent peptide quality and quantity across more than 3500 mass spectrometry runs.
This approach is especially beneficial for high-throughput applications, enabling rapid processing of large sample sets within clinically relevant time frames, thus paving the way for advanced proteomic studies with potential clinical applications. The HnB method represents a transformative step in proteomics, providing a streamlined, efficient, and scalable solution for researchers in this rapidly evolving field.
In the "Heat ‘n Beat" proteomics study, the NanoPhotometer N60 was used to measure the concentration of peptides via UV absorption at 280 nm, ensuring consistent loading onto the mass spectrometer for reliable analysis.
Visit www.implen.de to find out how the NanoPhotometer can improve your research.
Authors: Dylan Xavier, Natasha Lucas, Steven Williams, Jennifer Koh, Keith Ashman, Clare Loudon, Roger Reddel, Peter Hains, and Phillip Robinson.
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