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Cited by 38Protein purification and stabilityViral Infectious Diseases and Gene Expression in InsectsMonoclonal and Polyclonal Antibodies Research

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Elucidating the effects of pH shift on IgG1 monoclonal antibody acidic charge variant levels in Chinese hamster ovary cell cultures

Xie, Panpan, Niu, Huijie, Chen, Xinning, Zhang, Xintao, Miao, Shiwei, Deng, Xiancun, Liu, Xuping, Tan, Wen-Song, Zhou, Yan, Fan, Li·2016·Applied Microbiology and Biotechnology
DOI 10.1007/s00253-016-7749-4
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Abstract

Charge variants, especially acidic charge variants, in recombinant monoclonal antibodies are critical quality attributes, which can affect antibodies' properties in vitro and in vivo. Meanwhile, charge variants are cumulative effects of various post-translational modifications and chemical degradations on antibody. In this work, to investigate the effect of lowering culture pH in the stationary phase on acidic charge variant contents in fed-batch cultures and its mechanism, cell culture experiments in 2-L bioreactors were firstly performed to explore the changes in the charge distribution under the pH downshift condition using weak cation exchange chromatography. It is found that acidic charge variant contents were significantly decreased by pH downshift. Then, to reveal the mechanism by which the content of acidic charge variants is reduced under pH downshift condition, the variation of post-translational modifications and chemical degradations under the pH downshift condition was explored. Meanwhile, the structure of the acidic charge variants was characterized. Several analysis experiments including size exclusion chromatography, capillary electrophoresis-sodium dodecyl sulfate under non-reducing conditions, tryptic peptide map, and reduced antibody mass were applied in this study. The results show that the mechanism by which the content of acidic charge variants is reduced is that the contents of disulfide bond reduction, galactosylation, and asparagine deamination of the…

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Metadata

Authors
Xie, P., Niu, H., Chen, X., Zhang, X., Miao, S., Deng, X., Liu, X., Tan, W.-S., Zhou, Y., & Fan, L.
Affiliations
East China University of Science and Technology (CN)
Corresponding author
  • Zhou, Yan
  • Fan, Li
ORCID
Venue
Applied Microbiology and Biotechnology
Publisher
Springer Science and Business Media LLC
Published
2016-08-02
Volume
100
Issue
24
Pages
10343-10353
ISSN
  • 0175-7598 (ISSN-L)
  • 1432-0614
DOI
10.1007/s00253-016-7749-4
OpenAlex
W2488318721
Cited by
38
References
31
Citations by year
2026: 4 · 2025: 2 · 2024: 4 · 2023: 3 · 2022: 4 · 2021: 4 · 2020: 4 · 2019: 4 · 2018: 6 · 2017: 3
FWCI
2.25
Type
journal article
Language
en
Open access
closed
Licence
www.springer.com/tdm (unspecified)
Locations
  • Applied Microbiology and Biotechnology (publishedVersion)
  • PubMed (publishedVersion)
Funders
the National Natural Science Foundation of China (Nos. 21206040, Nos.21406066)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Life Sciences › Immunology and Microbiology › Immunology › Monoclonal and Polyclonal Antibodies Research
Keywords
IgG1 monoclonal antibody, culture medium pH, CHO cells, fed-batch fermentation, post-translational modification, disulfide bond reduction, galactosylation, Fc domain, antibody degradation, weak cation exchange chromatography
MeSH terms
Animals, Antibodies, Monoclonal, Cricetulus, Hydrogen-Ion Concentration, Immunoglobulin G, Protein Processing, Post-Translational, Recombinant Proteins, CHO Cells, Chemical Phenomena

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