Cited by 34Microfluidic and Capillary Electrophoresis ApplicationsMass Spectrometry Techniques and ApplicationsAdvanced Proteomics Techniques and Applications

Capillary zone electrophoresis tandem mass spectrometry detects low concentration host cell impurities in monoclonal antibodies

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Zhu, Guijie, Sun, Liangliang, Heidbrink‐Thompson, Jennifer, Kuntumalla, Srilatha, Lin, Hung‐yu, Larkin, Christopher J., McGivney, James B., Dovichi, Norman J.·2016·ELECTROPHORESIS

Abstract

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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Crossref · 10.1002/elps.201500301

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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.”

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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
Venue
ELECTROPHORESIS
Publisher
Wiley
Published
2016-02
Volume
37
Issue
4
Pages
616-622
ISSN
  • 0173-0835 (ISSN-L)
  • 1522-2683
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
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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