articleNature MethodsJan 16, 2025HYBRID OA

Challenging the Astral mass analyzer to quantify up to 5,300 proteins per single cell at unseen accuracy to uncover cellular heterogeneity

JAJulia A. BubisTNTabiwang N. ArreyEDEugen DamocBDBernard DelangheJSJana Slováková

Research Institute of Molecular Pathology · Thermo Fisher Scientific (Germany) · +5 more institutions

PubMed
Indexed incrossrefpubmed

Abstract

Despite significant advancements in sample preparation, instrumentation and data analysis, single-cell proteomics is currently limited by proteomic depth and quantitative performance. Here we demonstrate highly improved depth of proteome coverage as well as accuracy and precision for quantification of ultra-low input amounts. Using a tailored library, we identify up to 7,400 protein groups from as little as 250 pg of HeLa cell peptides at a throughput of 50 samples per day. Using a two-proteome mix, we check for optimal parameters of quantification and show that fold change differences of 2 can still be successfully determined at single-cell-level inputs. Eventually, we apply our workflow to A549 cells,…

Citation impact

83
total citations
FWCI
103.09
Percentile
100%
References
50
Citations per year

Authors

11

Topics & keywords

Topics
Keywords
  • Single-cell analysis
  • Computer science
  • Computational biology
  • Spectrum analyzer
  • Cell
  • Biophysics
  • Nanotechnology
  • Chemistry
No related works found for this paper.

Funding