Characterization of changes in the glycosylation pattern of recombinant proteins from BHK-21 cells due to different culture conditions.
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 (Glu100<==>Asn) generating a potential N-glycosylation recognition site (Asn100-Xxx-Thr/Ser) in addition to the natural O-glycosylation at position Thr3. 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 alpha 1-6 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.
Gawlitzek
M
M
Department for Cell Culture Techniques, Gesellschaft für Biotechnologische Forschung m.b.H., Braunschweig, Germany.
Valley
U
U
Nimtz
M
M
Wagner
R
R
Conradt
H S
HS
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry; D002455:Cell Division; D002478:Cells, Cultured; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D046508:Culture Techniques / Q000379:methods*; D006031:Glycosylation; D006801:Humans; D007376:Interleukin-2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007668:Kidney; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D002241:Carbohydrates; D016895:Culture Media, Serum-Free; D007376:Interleukin-2; D011134:Polysaccharides; D011994:Recombinant Proteins