Extracellular sialidase degrades sialic acid in recombinant human erythropoietin produced by an industrial Chinese hamster ovary cell strain / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1492-1499, 2012.
Article
in Zh
| WPRIM
| ID: wpr-233277
Responsible library:
WPRO
ABSTRACT
To investigate the N-glycosylation characteristics of recombinant human erythropoietin (rhEPO) produced by an industrial Chinese hamster ovary (CHO) cell line that is currently used in a large scale manufacturing process, we cultured this cell strain in static mode. The produced rhEPO in the culture supernatant was analyzed using isoelectric focusing (IEF) and Ricinus communis agglutinin-I (RCA-I) lectin precipitation. The lactate dehydrogenase (LDH) and sialidase activity in the serum-free supernatant were assayed as well. The analyses revealed that this cell strain could produce rhEPO with high sialic acid content, but during prolonged culture, cell viability decreased with time whilst the activity of sialidase present in the supernatant increased. The loss in rhEPO quality was due to a decrease in terminal sialic acid on the N-glycans, caused by sialidase degradation. The methods and findings in this paper serve as basis for further investigation of industrial production process.
Full text:
1
Index:
WPRIM
Main subject:
Recombinant Proteins
/
Genetic Engineering
/
Cricetulus
/
Erythropoietin
/
CHO Cells
/
Cell Culture Techniques
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N-Acetylneuraminic Acid
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Proteolysis
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Genetics
/
Metabolism
Limits:
Animals
/
Humans
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2012
Type:
Article