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An Automated Nanowell-Array Workflow for Quantitative Multiplexed Single-Cell Proteomics Sample Preparation at High
Claudia Ctortecka1, David Hartlmayr2, Anjali Seth3
1Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Vienna, Austria; Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Molecular & Cellular Proteomics : MCP
|October 15, 2023
Summary
The proteoCHIP streamlines single-cell proteomics sample preparation, enabling high-throughput, multiplexed analysis. This automated workflow enhances sensitivity and protein identification, facilitating biological discovery in single-cell studies.
Area of Science:
- Proteomics
- Cell Biology
- Analytical Chemistry
Background:
- Single-cell resolution reveals cellular heterogeneity in previously assumed homogenous populations.
- Advances in mass spectrometry instrumentation contrast with limitations in efficient single-cell sample preparation.
Purpose of the Study:
- To introduce the proteoCHIP, a universal, automated sample preparation method for multiplexed single-cell proteomics.
- To enhance sensitivity, throughput, and protein identification in single-cell proteomic analyses.
Main Methods:
- Automated single-cell isolation and picoliter dispensing using the cellenONE system.
- Multiplexed labeling up to 16-plex (TMT10-plex) with the proteoCHIP.
- Mass spectrometry acquisition strategies including wide precursor isolation windows (2 Da).
Main Results:
- Achieved high sensitivity and throughput for single-cell proteomics with the proteoCHIP.
- Identified approximately 1500 protein groups per TMT10-plex, with reduced need for carrier proteome.
- Identified an average of nearly 2000 proteins per TMT10-plex across 170 multiplexed single cells, distinguishing human cell types.
Conclusions:
- The proteoCHIP offers a versatile, automated, and sensitive solution for single-cell proteomics sample preparation.
- This workflow integrates efficient sample prep, advanced MS techniques, and data analysis for robust single-cell proteomic studies.
- The proteoCHIP approach is poised to advance biological applications of single-cell proteomics in research laboratories.
