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Related Experiment Video

Updated: Oct 8, 2025

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Quantitative Accuracy and Precision in Multiplexed Single-Cell Proteomics.

Claudia Ctortecka¹, Karel Stejskal^1,2,3, Gabriela Krššáková^1,2,3

¹Research Institute of Molecular Pathology (IMP), Vienna BioCenter (VBC), Campus Vienna Biocenter 1, 1030 Vienna, Austria.

Analytical Chemistry

|December 30, 2021

Summary

This summary is machine-generated.

Optimizing single-cell proteomics involves carefully managing carrier spike ratios. Reducing carrier abundance improves quantitative accuracy and data quality for trace sample analysis.

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Area of Science:

Proteomics
Biotechnology
Analytical Chemistry

Background:

Single-cell proteomics workflows have advanced, enabling detailed characterization of biological systems.
Multiplexed single-cell proteomics often uses abundant carriers to enhance precursor selection and identifications.
High carrier abundance may compromise quantitative accuracy despite increased total ion current.

Purpose of the Study:

To assess the impact of narrowly titrated carrier spikes (<20×) in single-cell proteomics.
To evaluate carrier elimination strategies for comparable sensitivity and superior accuracy.
To identify optimal multiplexing strategies and experimental designs for trace sample analysis.

Main Methods:

Focused analysis on narrowly titrated carrier spikes (<20×) in single-cell proteomics.
Evaluation of carrier ratio impact on measurement variability and data quality.
Benchmarking of multiplexing strategies, including DIA-TMT, for trace samples.

Main Results:

Subtle changes in carrier ratio significantly affect measurement variability.
Alternative multiplexing strategies were identified for assessing data quality.
Elevated replicate overlap and improved quantitative accuracy were achieved with DIA-TMT.

Conclusions:

Optimizing carrier spike ratios is crucial for accurate single-cell proteomics.
DIA-TMT offers a viable strategy for multiplexed proteomics of trace samples.
This work provides guidance for designing optimal single-cell proteomics experiments.