Combinatorial genome and protein engineering yields monoclonal antibodies with hypergalactosylation from CHO cells

Cheng Yu Chung, Qiong Wang, Shuang Yang, Sean A. Ponce, Brian J. Kirsch, Hui Zhang, Michael J. Betenbaugh

Research output: Contribution to journalArticlepeer-review


One of the key quality attributes of monoclonal antibodies is the glycan pattern and distribution. Two terminal galactose residues typically represent a small fraction of the total glycans from antibodies. However, antibodies with defined glycosylation properties including enhanced galactosylation have been shown to exhibit altered properties for these important biomedical modalities. In this study, the disruption of two α-2,3 sialyltransferases (ST3GAL4 and ST3GAL6) from Chinese Hamster Ovary (CHO) cells was combined with protein engineering of the Fc region to generate an IgG containing 80% bigalactosylated and fucosylated (G2F) glycoforms. Expression of the same single amino acid mutant (F241A) IgG in CHO cells with a triple gene knockout of fucosyltransferase (FUT8) plus ST3GAL4 and ST3GAL6 lowered the galactosylation glycoprofile to 65% bigalactosylated G2 glycans. However, overexpression of IgGs with four amino acid substitutions recovered the G2 glycoform composition approximately 80%. Combining genome and protein engineering in CHO cells will provide a new antibody production platform that enables biotechnologists to generate glycoforms standards for specific biomedical and biotechnology applications.

Original languageEnglish (US)
Pages (from-to)2848-2856
Number of pages9
JournalBiotechnology and bioengineering
Issue number12
StatePublished - Dec 2017


  • CRISPR/Cas 9 genome editing
  • Chinese hamster ovary cells
  • antibody engineering
  • glycoengineering

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


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