Skip to content
Challenging the Astral mass analyzer – going beyond 5200 proteins per single-cell at unseen quantitative accuracy to study cellular heterogeneity.

In a study focused on single-cell proteomics, researchers aimed to overcome limitations related to proteomic depth and quantitative performance. They achieved this by using a combination of a zero dead-end volume chromatographic column and the Thermo Scientific Orbitrap Astral mass spectrometer in data-independent acquisition (DIA) mode. This approach allowed them to achieve an unprecedented depth of proteome coverage and accurate quantification, even at ultra-low input amounts.

Using a tailored library, the researchers identified up to 7,400 protein groups from as little as 250 pg of HeLa cells at a throughput of 50 samples per day. They also benchmarked different data analysis strategies, demonstrating that false discovery rates (FDR) on the protein level can be maintained at 1%. Additionally, they successfully determined fold change differences of 2 at single-cell input levels.

The workflow was applied to A549 cells, resulting in the identification of up to 5,200 protein groups from a single cell, enabling the study of cellular heterogeneity and dependencies between cell size and cell-cycle phase. The researchers also used the workflow to distinguish between in vitro analogs of two human blastocyst lineages.

Overall, this study represents a significant advancement in single-cell proteomics, offering improved sensitivity, reproducibility, and quantitative accuracy, making it a valuable tool for biologists to complement single-cell genomics data.

The study utilized an Aurora Ultimate TS 25 cm nanoflow UHPLC, enabling the achievement of unprecedented proteome coverage and accuracy in quantifying ultra-low input amounts in single-cell proteomics.


BioRxiv – Feb 1, 2024

Authors

J. A. Bubis; T. N. Arrey; E Damoc; B. Delanghe; J Slovakova; T M. Sommer; H. Kagawa,; P Pichler; N. Rivron, K. Mechtler, M. Matzinger

Title

Challenging the Astral mass analyzer – going beyond 5200 proteins per single-cell at unseen quantitative accuracy to study cellular heterogeneity.

Share:

Related publications & news

World Immunization Week starts today! From the 24 to the 30 April 2024...
Members of the IonOpticks team celebrate receiving the award at its manufacturing facility in Fitzroy, Melbourne.
Melbourne, April 22 2024: IonOpticks has proudly been awarded a Silver Winn...
This groundbreaking study introduces a novel deep learning-based approach c...
In a groundbreaking study, researchers used Single Cell ProtEomics to delve...
Leduc et. al have developed nPOP, a revolutionary method for single-cell pr...
Presented here is the latest breakthrough in immunopeptidomics by Wahle et ...
In a groundbreaking study by Gurung et. al, researchers expanded the repert...
Researchers have made significant strides in understanding the sexual repro...
Previous
Next