Glutamine-free mammalian expression of recombinant glycoproteins with uniform isotope labeling: an application for NMR analysis of pharmaceutically relevant Fc glycoforms of human immunoglobulin G1

• DOI: 10.1007/s10858-021-00387-5
• Publication Date: 2022-01-03T00:03:15Z
• Authors: Saeko Yanaka; Hirokazu Yagi; Rina Yogo; Masayoshi Onitsuka; Koichi Kato
• Keywords: Mammals; 0303 health sciences; 03 medical and health sciences; Glutamine; Isotope Labeling; Animals; Humans; Immunoglobulins; Nuclear Magnetic Resonance, Biomolecular; Recombinant Proteins; Glycoproteins; 3. Good health

Mammalian cells are widely used for producing recombinant glycoproteins of pharmaceutical interest. However, a major drawback of using mammalian cells is the high production costs associated with uniformly isotope-labeled glycoproteins due to the large quantity of labeled L-glutamine required for their growth. To address this problem, we developed a cost-saving method for uniform isotope labeling by cultivating the mammalian cells under glutamine-free conditions, which was achieved by co-expression of glutamine synthase. We demonstrate the utility of this approach using fucosylated and non-fucosylated Fc glycoforms of human immunoglobulin G1.