Interlaboratory study to evaluate the robustness of capillary electrophoresis-mass spectrometry for peptide mapping

Christian Wenz*, Coral Barbas, Ángeles Lõpez-Gonzálvez, Antonia Garcia, Fernando Benavente, Victoria Sanz-Nebot, Tim Blanc, Gordon Freckleton, Philip Britz-McKibbin, Meera Shanmuganathan, Francois de l'Escaille, Johann Far, Rob Haselberg, Sean Huang, Carolin Huhn, Martin Pattky, David Michels, Si Mou, Feng Yang, Christian NeusuessNora Tromsdorf, Edward E. K. Baidoo, Jay D. Keasling, SungAe Suhr Park

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review


A collaborative study on the robustness and portability of a capillary electrophoresis-mass spectrometry method for peptide mapping was performed by an international team, consisting of 13 independent laboratories from academia and industry. All participants used the same batch of samples, reagents and coated capillaries to run their assays, whereas they utilized the capillary electrophoresis-mass spectrometry equipment available in their laboratories. The equipment used varied in model, type and instrument manufacturer. Furthermore, different types of sheath-flow capillary electrophoresis-mass spectrometry interfaces were used. Migration time, peak height and peak area of ten representative target peptides of trypsin-digested bovine serum albumin were determined by every laboratory on two consecutive days. The data were critically evaluated to identify outliers and final values for means, repeatability (precision within a laboratory) and reproducibility (precision between laboratories) were established. For relative migration time the repeatability was between 0.05 and 0.18% RSD and the reproducibility between 0.14 and 1.3% RSD. For relative peak area repeatability and reproducibility values obtained were 3-12 and 9-29% RSD, respectively. These results demonstrate that capillary electrophoresis-mass spectrometry is robust enough to allow a method transfer across multiple laboratories and should promote a more widespread use of peptide mapping and other capillary electrophoresis-mass spectrometry applications in biopharmaceutical analysis and related fields.

Original languageEnglish
Pages (from-to)3262-3270
Number of pages9
JournalJournal of Separation Science
Issue number18
Publication statusPublished - 1 Sept 2015


  • Precision
  • Repeatability
  • Reproducibility
  • System suitability test
  • Tryptic digest


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