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Function characterization of a glyco-engineered anti-EGFR monoclonal antibody cetuximab in vitro.

Acta pharmacologica Sinica (2014-09-30)
Chang-hong Yi, Can-ping Ruan, Hao Wang, Xin-yun Xu, Yun-peng Zhao, Meng Fang, Jun Ji, Xing Gu, Chun-fang Gao

To evaluate the biochemical features and activities of a glyco-engineered form of the anti-human epidermal growth factor receptor monoclonal antibody (EGFR mAb) cetuximab in vitro. The genes encoding the Chinese hamster bisecting glycosylation enzyme (GnTIII) and anti-human EGFR mAb were cloned and coexpressed in CHO DG44 cells. The bisecting-glycosylated recombinant EGFR mAb (bisec-EGFR mAb) produced by these cells was characterized with regard to its glycan profile, antiproliferative activity, Fc receptor binding affinity and cell lysis capability. The content of galactose-α-1,3-galactose (α-Gal) in the bisec-EGFR mAb was measured using HPAEC-PAD. The bisec-EGFR mAb had a higher content of bisecting N-acetylglucosamine residues. Compared to the wild type EGFR mAb, the bisec-EGFR mAb exhibited 3-fold higher cell lysis capability in the antibody-dependent cellular cytotoxicity assay, and 1.36-fold higher antiproliferative activity against the human epidermoid carcinoma line A431. Furthermore, the bisec-EGFR mAb had a higher binding affinity for human FcγRIa and FcγRIIIa-158F than the wild type EGFR mAb. Moreover, α-Gal, which was responsible for cetuximab-induced hypersensitivity reactions, was not detected in the bisec-EGFR mAb. The glyco-engineered EGFR mAb with more bisecting modifications and lower α-Gal content than the approved therapeutic antibody Erbitux shows improved functionality in vitro, and requires in vivo validations.

Product Number
Product Description

Galactose, European Pharmacopoeia (EP) Reference Standard
D-(+)-Galactose, meets analytical specification of Ph. Eur., BP
D-(+)-Galactose, suitable for microbiology, ≥99.0%
D-(+)-Galactose, ≥98%
D-(+)-Galactose, powder, anhydrous, BioReagent, suitable for cell culture, suitable for insect cell culture
D-(+)-Galactose, ≥99%
Galactose, Pharmaceutical Secondary Standard; Certified Reference Material
Galactose, plant-derived
Galactose, United States Pharmacopeia (USP) Reference Standard
D-(+)-Galactose, BioXtra, pH 5.0-7.0 (20 °C, 1 M in H2O), ≥99%