An Improved Boosting to Amplify Signal with Isobaric Labeling (iBASIL) Strategy for Precise Quantitative Single-cell Proteomics.

Molecular & Cellular Proteomics : MCP
Chia-Feng TsaiTao Liu

Abstract

Mass spectrometry (MS)-based proteomics has great potential for overcoming the limitations of antibody-based immunoassays for antibody-independent, comprehensive, and quantitative proteomic analysis of single cells. Indeed, recent advances in nanoscale sample preparation have enabled effective processing of single cells. In particular, the concept of using boosting/carrier channels in isobaric labeling to increase the sensitivity in MS detection has also been increasingly used for quantitative proteomic analysis of small-sized samples including single cells. However, the full potential of such boosting/carrier approaches has not been significantly explored, nor has the resulting quantitation quality been carefully evaluated. Herein, we have further evaluated and optimized our recent boosting to amplify signal with isobaric labeling (BASIL) approach, originally developed for quantifying phosphorylation in small number of cells, for highly effective analysis of proteins in single cells. This improved BASIL (iBASIL) approach enables reliable quantitative single-cell proteomics analysis with greater proteome coverage by carefully controlling the boosting-to-sample ratio (e.g. in general <100×) and optimizing MS automatic gain contr...Continue Reading

References

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Citations

Aug 28, 2020·Molecular & Cellular Proteomics : MCP·Ryan T Kelly
Dec 9, 2020·Nature Methods·Tommy K CheungChristopher M Rose
Dec 23, 2020·The FEBS Journal·Indranil PaulAndrew Emili
Dec 16, 2020·Proteomics·Timothy D Veenstra
Jan 28, 2021·Critical Reviews in Analytical Chemistry·Negin Fasih RamandiHassan Y Aboul-Enein
Jun 25, 2020·Molecular & Cellular Proteomics : MCP·Daniel G Delafield, Lingjun Li
Feb 26, 2021·Analytical Methods : Advancing Methods and Applications·Zhichang Yang, Liangliang Sun
Jul 1, 2020·Current Opinion in Chemical Biology·Nikolai Slavov

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Datasets Mentioned

BETA
PXD016057
PXD017626
JPST000690
JPST000723

Methods Mentioned

BETA
protein assay
chip
acetylation
PCA

Software Mentioned

iBASIL
BASIL
Perseus
MaxQuant

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