Effect of N-Acetylcystein on butyrate-treated Chinese hamster ovary cells to improve the production of recombinant human interferon-beta-1a.
Sodium butyrate (NaBu) is used as a productivity enhancer for the production of therapeutic recombinant proteins in Chinese hamster ovary (CHO) cells. However, NaBu is well-known for having a cytotoxic effect, thereby inducing apoptosis. As an endeavor to reduce this defect, we studied 11 antioxidants known for inhibiting apoptosis, according to a Plackett-Burman statistical design on CHO cells producing recombinant interferon-beta-1a (IFN-beta). None of the antioxidants that we tested were as effective as N-acetylcystein (NAC) from the point of view of maintaining long-term survival of CHO cells and increasing the production of IFN-beta. In 7.5-L perfusion bioreactor cultures, the addition of NaBu and NAC elongated the culture period to almost 200 h throughout production phase and increased the production yield by 2-fold compared to control cultures containing only NaBu. Glycosylation patterns of produced IFN-beta at each run were also compared in IEF analysis. IEF profiles of where NaBu and NAC were added showed to be more isoforms with a lower pI than those of the control run. The sialic acid content was also increased by 17.7% according to HPLC analysis. Taken together, the data obtained demonstrate that the addition of NAC has positive effects on the elongation of the culture period, improving the production and increasing the sialylation of IFN-beta in NaBu-treated CHO cells.
Oh
Han Kyu
HK
Life Science Research Team, R&D Center, Samsung Fine Chemicals Co., Ltd., Rm. 6113, College of Medicine, SKKU 300, Chunchun-Dong, Jangan-Gu, Suwon, Kyunggi-Do, Korea.
So
Moon Kyoung
MK
Yang
Jihye
J
Yoon
Ho Chul
HC
Ahn
Ji Soo
JS
Lee
Jong Min
JM
Kim
Ji Tai
JT
Yoo
Ji Uk
JU
Byun
Tae Ho
TH
D000111:Acetylcysteine / Q000494:pharmacology*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D019149:Bioreactors; D002087:Butyrates / Q000494:pharmacology*; D016466:CHO Cells / Q000187:drug effects; D053148:Caspase 3; D020169:Caspases / Q000187:drug effects / Q000378:metabolism; D018929:Cell Culture Techniques / Q000295:instrumentation; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D045304:Cytochromes c / Q000187:drug effects / Q000378:metabolism; D053938:DNA Fragmentation; D004686:Encephalomyocarditis virus / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007370:Interferon Type I / Q000187:drug effects / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D000068556:Interferon beta-1a; D016899:Interferon-beta / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D011994:Recombinant Proteins
D000975:Antioxidants; D002087:Butyrates; D007370:Interferon Type I; D011994:Recombinant Proteins; D016899:Interferon-beta; D045304:Cytochromes c; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D005978:Glutathione; D019158:N-Acetylneuraminic Acid; D000111:Acetylcysteine; D000068556:Interferon beta-1a