Characterization of changes in the glycosylation pattern of recombinant proteins from BHK-21 cells due to different culture conditions

• Published in: Journal of biotechnology 1995-09, Vol.42 (2), p.117-131
• Main Authors: Gawlitzek, Martin, Valley, Ulrich, Nimtz, Manfred, Wagner, Roland, Conradt, Harald S.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animal cells; Animals; Biological and medical sciences; Biotechnology; Blotting, Western; Carbohydrate pattern; Carbohydrate Sequence; Carbohydrates - chemistry; Cell Division; Cells, Cultured; Continuous perfused bioreactor; Cricetinae; Culture Media, Serum-Free; Culture Techniques - methods; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Glycoprotein; Glycosylation; Humans; Interleukin-2 - chemistry; Interleukin-2 - genetics; Interleukin-2 - metabolism; Kidney; Methods. Procedures. Technologies; Models, Molecular; Molecular Sequence Data; Polysaccharides - biosynthesis; Protein free medium; Recombinant mammalian cell; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/0168-1656(95)00065-X

The N-glycosylation patterns of a genetically engineered human interleukin-2 variant glycoprotein (IL-Mu6), produced by BHK-21 cells from long-term suspension and microcarrier cultures in the presence and absence of fetal calf serum were compared. IL-Mu6 was used as a model protein in studying the effect of different controlled cell culture conditions on the expression of N-glycans in recombinant glycoproteins. IL-Mu6 contains a single amino acid substitution (Glu 100 ↔ Asn) generating a potential N-glycosylation recognition site (Asn 100-Xxx-Thr/Ser) in addition to the natural O-glycosylation at position Thr 3. Parallel cell cultivations were carried out in two continuously perfused 2.5-liter stirred bioreactors under defined culture conditions. Major differences were found in the glycoprotein products obtained during these different cultivation conditions. Serum-free cultures resulted in a higher level of terminal sialylation and proximal α1–6 fucosylation. The ratio of O- to N-glycans as well as the amount of nonglycosylated product and the antennarity of N-linked carbohydrates in the model protein exhibited major differences depending on the presence or absence of serum, the condition of growth and the cultivation procedure.