Effect of glucose feeding on the glycosylation quality of antibody produced by a human cell line, F2N78, in fed-batch culture
| Autor: | Myung Sam Cho, Yeon Jung Kim, Jong Moon Cho, Gyun Min Lee, Jin Seok Seo, Eric Baek, Byung Sup Min |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Glycosylation
Carbohydrate metabolism Applied Microbiology and Biotechnology Antibodies Cell Line chemistry.chemical_compound Western blot N-linked glycosylation Batch Cell Culture Techniques medicine Humans biology medicine.diagnostic_test General Medicine Recombinant Proteins Culture Media Fed-batch culture Glucose chemistry Biochemistry Cell culture biology.protein Antibody Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology. 98:3509-3515 |
| ISSN: | 1432-0614 0175-7598 |
| Popis: | The human cell line rF2N78 produces an antibody with a high galactosylation ratio which resembles human IgG. However, it has been observed that the aglycosylated antibody starts to appear when glucose is depleted. To determine whether glucose depletion is a main cause for aglycosylation of the antibody, fed-batch cultures of rF2N78 cells were performed using different feeding cocktails (glucose only, nutrient feeding cocktail without glucose, and nutrient feeding cocktail with glucose). In the fed-batch culture with nutrient feeding cocktail without glucose, aglycosylated antibody was produced in a later phase of culture, when glucose was depleted. Approximately 44 % of antibodies produced were aglycosylated at the end of culture. In contrast, aglycosylated antibody was not produced in cultures with glucose feeding. The expression levels of oligosaccharyl transferases determined by Western blot analysis were similar among the cultures, suggesting that aglycosylation of the antibody was not due to altered expression of oligosaccharyl transferases under glucose-deficient conditions. Thus, it is likely that glucose deficiency led to insufficiency of the precursor for glycosylation and induced aglycosylation of the antibody. Taken together, glucose feeding in fed-batch cultures successfully prevented occurrence of aglycosylated antibody during the cultures, confirming that glucose depletion is a main cause for aglycosylation of antibody. |
| Databáze: | OpenAIRE |
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