Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

High-Throughput Quantification of Spike Protein Expression by LC-MS for mRNA-LNP Evaluation and Immune Response Profiling.

Analytical chemistry·2026
Same author

Characterisation of a cohort of opportunistically recruited patients with COVID-19 and approaches to patient stratification.

BMC infectious diseases·2026
Same author

A Novel Compact Multi-Reflecting Time-of-Flight Mass Spectrometer.

Journal of the American Society for Mass Spectrometry·2026
Same author

Ion Mobility Separation of Isomeric Acyl-lysine Marks in Peptides.

Analytical chemistry·2026
Same author

Carotid Artery Stenting.

Journal of the Society for Cardiovascular Angiography & Interventions·2025
Same author

Enhanced Declustering Enables Native Top-Down Analysis of Membrane Protein Complexes using Ion-Mobility Time-Aligned Fragmentation.

Journal of the American Society for Mass Spectrometry·2024

Related Experiment Video

Updated: May 20, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

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

Published on: November 15, 2017

Using ion purity scores for enhancing quantitative accuracy and precision in complex proteomics samples.

Scott J Geromanos1, Chris Hughes, Steven Ciavarini

  • 1Waters Corporation, Milford, MA 01757-3696, USA. scott_geromanos@waters.com

Analytical and Bioanalytical Chemistry
|July 20, 2012
PubMed
Summary

Ion interference significantly impacts quantitative proteomics. Increasing analytical selectivity, like mass resolution and ion mobility, reduces interference, leading to more accurate peptide and protein quantification in LC-MS experiments.

More Related Videos

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry
11:12

Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry

Published on: February 7, 2022

Related Experiment Videos

Last Updated: May 20, 2026

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

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

Published on: November 15, 2017

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling
09:35

Resolving Affinity Purified Protein Complexes by Blue Native PAGE and Protein Correlation Profiling

Published on: April 1, 2017

Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry
11:12

Organic Solvent-Based Protein Precipitation for Robust Proteome Purification Ahead of Mass Spectrometry

Published on: February 7, 2022

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Mass Spectrometry

Background:

  • Accurate quantification of peptides and proteins in complex samples is crucial.
  • Ion interference events can compromise the precision of calculated ion areas.
  • Existing methods may not fully account for ion detection uniqueness.

Purpose of the Study:

  • To quantify ion interference events in quantitative proteomics using Stable Isotope Labeling by Amino acids in Cell culture (SILAC) experiments.
  • To evaluate the impact of analytical selectivity, including mass resolving power and ion mobility, on ion interference rates.
  • To develop ion purity scores for assessing the accuracy and precision of quantitative results.

Main Methods:

  • Comparison of "light" and "heavy" labeled ion detections in binary SILAC experiments based on m/z and retention time.
  • Determination of ion interference rates using Liquid Chromatography-Mass Spectrometry (LC-MS) at varying mass resolving powers (20K, 40K) with and without ion mobility.
  • Calculation of effective resolution, mass, chromatographic, and drift purity scores based on instrument performance and ion detection parameters.

Main Results:

  • A decrease in ion interference events was observed with increased analytical selectivity.
  • Ion purity scores can be calculated using mass resolving power and ion detection data (m/z, $\Delta$ mass at half-height).
  • Error in calculated peak area is estimated by comparing measured widths to experimental medians.

Conclusions:

  • Ion interference is a common issue in quantitative proteomics LC-MS.
  • Ion purity filters can effectively mitigate ion interference.
  • Implementing ion purity assessment leads to more accurate and precise quantification across a wider dynamic range.