Glycoengineering Chinese hamster ovary cells: a short history

Donini, Roberto; Haslam, Stuart M.; Kontoravdi, Cleo

doi:10.1042/bst20200840

Cited by 16 publications

(10 citation statements)

References 151 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the literature reports that low levels of terminal α1,3 Gal epitopes are possible in CHO cells, they are generally very low or almost absent in many CHO cell lines, including the one used in this work. 25 , 32 , 33 …”

Section: Discussionmentioning

confidence: 99%

A glyco-engineering approach for site-specific conjugation to Fab glycans

Jaramillo

Sulea

Durocher

et al. 2022

mAbs

View full text Add to dashboard Cite

Effective processes for synthesizing antibody-drug conjugates (ADCs) require: 1) site-specific incorporation of the payload to avoid interference with binding to the target epitope, 2) optimal drug/antibody ratio to achieve sufficient potency while avoiding aggregation or solubility problems, and 3) a homogeneous product to facilitate approval by regulatory agencies. In conventional ADCs, the drug molecules are chemically attached randomly to antibody surface residues (typically Lys or Cys), which can interfere with epitope binding and targeting, and lead to overall product heterogeneity, long-term colloidal instability and unfavorable pharmacokinetics. Here, we present a more controlled process for generating ADCs where drug is specifically conjugated to only Fab N -linked glycans in a narrow ratio range through functionalized sialic acids. Using a bacterial sialytransferase, we incorporated N -azidoacetylneuraminic acid (Neu5NAz) into the Fab glycan of cetuximab. Since only about 20% of human IgG1 have a Fab glycan, we extended the application of this approach by using molecular modeling to introduce N -glycosylation sites in the Fab constant region of other therapeutic monoclonal antibodies. We used trastuzumab as a model for the incorporation of Neu5NAz in the novel Fab glycans that we designed. ADCs were generated by clicking the incorporated Neu5NAz with monomethyl auristatin E (MMAE) attached to a self-immolative linker terminated with dibenzocyclooctyne (DBCO). Through this process, we obtained cetuximab-MMAE and trastuzumab-MMAE with drug/antibody ratios in the range of 1.3 to 2.5. We confirmed that these ADCs still bind their targets efficiently and are as potent in cytotoxicity assays as control ADCs obtained by standard conjugation protocols. The site-directed conjugation to Fab glycans has the additional benefit of avoiding potential interference with effector functions that depend on Fc glycan structure.

show abstract

Section: Discussionmentioning

confidence: 99%

A glyco-engineering approach for site-specific conjugation to Fab glycans

Jaramillo

Sulea

Durocher

et al. 2022

mAbs

View full text Add to dashboard Cite

show abstract

“…Recent breakthroughs in gene editing have revolutionized glycoengineering in mammalian cells and led to improved designs of therapeutic proteins [ 1 , 6 , 12 ]. The consistent production of safer and potentially more efficacious biotherapeutics have been the primary goals for these efforts.…”

Section: Glycoengineering As a Continued Theme For Biotherapeutics Applicationsmentioning

confidence: 99%

“…Therapeutic proteins, such as antibodies and recombinant fusion proteins, are glycoproteins in which glycan modifications are often considered critical quality attributes and can be engineered for therapeutic efficacy and safety improvements (according to several reviews [ 6 , 10 , 11 , 12 , 13 ]). With a global view on the human glycome being established and a deeper understanding on glycosylation pathways, new opportunities in harnessing human protein glycosylation functions are emerging ( Figure 1 ).…”

Section: Introductionmentioning

confidence: 99%

New Opportunities in Glycan Engineering for Therapeutic Proteins

et al. 2022

View full text Add to dashboard Cite

Glycans as sugar polymers are important metabolic, structural, and physiological regulators for cellular and biological functions. They are often classified as critical quality attributes to antibodies and recombinant fusion proteins, given their impacts on the efficacy and safety of biologics drugs. Recent reports on the conjugates of N-acetyl-galactosamine and mannose-6-phosphate for lysosomal degradation, Fab glycans for antibody diversification, as well as sialylation therapeutic modulations and O-linked applications, have been fueling the continued interest in glycoengineering. The current advancements of the human glycome and the development of a comprehensive network in glycosylation pathways have presented new opportunities in designing next-generation therapeutic proteins.

show abstract

“…Considering the fact that the gene of α‐1,3 galactosidase was silenced or low expressed in Chinese hamster ovary (CHO) cell line, which would avoid the α‐Gal modification (Donini et al, 2021), a recombinant anti‐EGFR antibody (code: AB01) was expressed by CHO cell line. In this study, the structure properties, affinity, antiproliferative effect and endocytosis behavior have been examined to evaluate the difference between mammalian cell‐expressed and insect cell‐expressed anti‐EGFR mAbs.…”

Section: Introductionmentioning

confidence: 99%

Structure and activity of ananti‐epidermal growth factor receptor antibody without galactose‐α‐1,3‐galactose residues

Ren

Gong

et al. 2021

Drug Dev Res

View full text Add to dashboard Cite

Epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein, which has been considered as one of the key targets for cancer therapy. However, currently approved therapeutic anti‐EGFR antibody may cause the hypersensitivity reaction induced by galactose‐α‐1,3‐galactose (α‐Gal) structure, which is inevitable in insect cell expression system. In this study, the Chinese hamster ovary cell line was used to produce a monoclonal antibody containing simplified glycosylation patterns (code: AB01). And cetuximab was used as a control. The two antibodies were highly similar in molecular weight, secondary structure, binding affinity and endocytosis behavior, whereas the glycotypes are extremely distinct. The flow cytometry assay suggested that AB01 induced cell cycle arrest in G1, thus inhibit cell proliferation. Moreover, both cetuximab and AB01 showed similar sensitivity for all tested cell lines in this research. In conclusion, AB01 could be a potential anti‐EGFR drug candidate for cancer therapy.

show abstract

Glycoengineering Chinese hamster ovary cells: a short history

Cited by 16 publications

References 151 publications

A glyco-engineering approach for site-specific conjugation to Fab glycans

A glyco-engineering approach for site-specific conjugation to Fab glycans

New Opportunities in Glycan Engineering for Therapeutic Proteins

Structure and activity of ananti‐epidermal growth factor receptor antibody without galactose‐α‐1,3‐galactose residues

Contact Info

Product

Resources

About