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

Abstract

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.

Evaluation of sample injection precision in respect to sensitivity in capillary electrophoresis using various injection modes

“…In turn, the preconcentration strategies might be considered for biological samples analysis whenever the required sensitivity cannot be reached with standard approach and for which the acceptance criteria are less stringent (CV < 20% in European Medicines Agency). Despite the precision of CE in coupling with MS (which detector is known to feature inferior repeatability in comparison to UV detector) has recently been proved satisfactory in interlaboratory study [22], the experimental results lead to the conclusion that significant improvements in CE instrumentation are of highest demand. Introduction of more precise injectors (e.g., valve based) to the commercial devices might significantly contribute to the applicability of CE and might solve a number of problems of this powerful technique.…”

Recent advances in CE‐MS coupling: Instrumentation, methodology, and applications

“…Although there is a number of crucial issues that are to be solved, the current progress of CE‐MS has enabled to realize many unique analyses and construct reliable, commercially available instrumentation. A very good example can be interlaboratory test done in 13 laboratories of sheath‐flow interfacing reaching (on different interface model type) RSD of migration times 0.14–1.3% . However, in recent years electrokinetically pumped sheath‐flow ESI interface with nL/min flow rate and sheathless interface with porous tip were commercialized, significantly contributing to better robustness of analyses.…”

“…The analyte/sheathliquid mass flow rates ratio determines analyte dilution and influences the sensitivity of the analysis. The sheath‐flow interfaces with several μL/min flow rates represent very powerful tools for extensive clinical and biological studies due to a reasonable robustness and reproducible results at interlaboratory level . However, the use of low sheath‐flow interfacing with electrokinetically pumped sheathliquid is on its rise nowadays.…”

“…The interlaboratory robustness of CE‐MS for peptide mapping was studied by several groups. . The most intensive is the effort in the investigation of tumor markers.…”

Revealing the potential of capillary electrophoresis/mass spectrometry: the tipping point

“…In particular, a similar CE/MS/MS methodology could be used to investigate the similarity between different therapeutic monoclonal antibodies (mAbs) and their respective biosimilar candidates which emphasizes the outstanding level of characterization and robustness of the data generated . Also, CE/MS could be validated through inter‐laboratory studies showing the transferability and robustness of this instrumental approach …”