Capillary zone electrophoresis tandem mass spectrometry detects low concentration host cell impurities in monoclonal antibodies
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We have evaluated CZE‐ESI‐MS/MS for detection of trace amounts of host cell protein impurities in recombinant therapeutics. Compared to previously published procedures, we have optimized the buffer pH used in the formation of a pH junction to increase injection volume. We also prepared a 5‐point calibration curve by spiking 12 standard proteins into a solution of a human mAb. A custom CZE‐MS/MS system was used to analyze the tryptic digest of this mixture without depletion of the antibody. CZE generated a ∼70‐min separation window (∼90‐min total analysis duration) and ∼300‐peak capacity. We also analyzed the sample using ultra‐performance LC‐MS/MS. CZE‐MS/MS generated approximately five times higher base peak intensity and more peptide identifications for low‐level spiked proteins. Both methods detected all proteins spiked at ∼100 ppm level with respect to the antibody.
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CZE has proven valuable for protein identification via bottom-up proteomics, where ultrasensitive and fast analysis can be obtained.
“CZE has also proven to be a valuable tool for protein identification via bottom-up proteomics where ultrasensitive and fast analysis can be obtained72–74.”
From Recent trends of capillary electrophoresis‐mass spectrometry in proteomics research (2019) · cites this paper as [74]
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Recent trends of capillary electrophoresis‐mass spectrometry in proteomics research
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- Authors
- Zhu, G., Sun, L., Heidbrink‐Thompson, J., Kuntumalla, S., Lin, H., Larkin, C. J., McGivney, J. B., & Dovichi, N. J.
- Affiliations
- University of Notre Dame (US)
- Department of Analytical Biotechnology MedImmune Gaithersburg MD USA
- Department of Chemistry and Biochemistry University of Notre Dame Notre Dame IN USA
- ORCID
- Sun, Liangliang
- Venue
- ELECTROPHORESIS
- Publisher
- Wiley
- Published
- 2016-02
- Volume
- 37
- Issue
- 4
- Pages
- 616-622
- ISSN
- 0173-0835 (ISSN-L)
- 1522-2683
- PMC
- PMC4816075
- OpenAlex
- W1953741266
- Cited by
- 34
- References
- 39
- Citations by year
- 2025: 1 · 2022: 5 · 2021: 4 · 2020: 4 · 2019: 2 · 2018: 2 · 2017: 5 · 2016: 9 · 2015: 1 · 2013: 1
- FWCI
- 2.82
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Electrophoresis (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of General Medical Sciences (R01GM096767)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- capillary zone electrophoresis, tandem mass spectrometry, monoclonal antibodies, trace impurity detection, injection volume, tryptic digest, peptide identification
- MeSH terms
- Antibodies, Monoclonal, Drug Contamination, Escherichia coli, Humans, Electrophoresis, Capillary, Spectrometry, Mass, Electrospray Ionization, Tandem Mass Spectrometry, Chromatography, Reverse-Phase
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