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Direct quality control of glycoengineered erythropoietin variants

Author

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  • Tomislav Čaval

    (University of Utrecht
    Netherlands Proteomics Center)

  • Weihua Tian

    (University of Copenhagen)

  • Zhang Yang

    (University of Copenhagen)

  • Henrik Clausen

    (University of Copenhagen)

  • Albert J. R. Heck

    (University of Utrecht
    Netherlands Proteomics Center)

Abstract

Recombinant production of glycoprotein therapeutics like erythropoietin (EPO) in mammalian CHO cells rely on the heterogeneous N-glycosylation capacity of the cell. Recently, approaches for engineering the glycosylation capacities of mammalian cells for custom designed glycoforms have been developed. With these opportunities there is an increasing need for fast, sensitive, and global analysis of the glycoproteoform landscape produced to evaluate homogeneity and consistency. Here we use high-resolution native mass spectrometry to measure the glycoproteoform profile of 24 glycoengineered variants of EPO. Based on the unique mass and intensity profiles of each variant, we classify them according to similarities in glycosylation profiles. The classification distinguishes EPO variants with varying levels of glycan branchingand sialylation, which are crucial parameters in biotherapeutic efficacy. We propose that our methods could be of great benefit in the characterization of other glycosylated biopharmaceuticals, ranging from the initial clonal selection to batch-to-batch controls, and the assessment of similarity between biosimilar/biobetter products.

Suggested Citation

  • Tomislav Čaval & Weihua Tian & Zhang Yang & Henrik Clausen & Albert J. R. Heck, 2018. "Direct quality control of glycoengineered erythropoietin variants," Nature Communications, Nature, vol. 9(1), pages 1-8, December.
  • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-05536-3
    DOI: 10.1038/s41467-018-05536-3
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    Cited by:

    1. Luis F. Schachner & Christopher Mullen & Wilson Phung & Joshua D. Hinkle & Michelle Irwin Beardsley & Tracy Bentley & Peter Day & Christina Tsai & Siddharth Sukumaran & Tomasz Baginski & Danielle DiCa, 2024. "Exposing the molecular heterogeneity of glycosylated biotherapeutics," Nature Communications, Nature, vol. 15(1), pages 1-13, December.

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