کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
4753624 1416992 2017 11 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Cell culture media supplemented with raffinose reproducibly enhances high mannose glycan formation
ترجمه فارسی عنوان
محتویات سلول های کشت سلولی حاوی رافینوز به طور مجدد باعث افزایش تشکیل گلیکان منوژن می شود
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی


- Raffinose is presented as a novel cell culture medium supplement to promote high mannose N-glycans in CHO fed-batch cultures.
- Two recombinant CHO cell lines were tested in high throughput cell culture systems and lab-scale bioreactors.
- Up to six-fold increases of high mannose species were observed with increasing raffinose concentrations in the medium.
- Our results show the potential of medium supplementation to alter glycosylation patterns of recombinant proteins.

Glycosylation plays a pivotal role in pharmacokinetics and protein physiochemical characteristics. In particular, effector functions including antibody-dependent cell-mediated cytotoxicity (ADCC) can be desired, and it has been described that high-mannose species exhibited enhanced ADCC. In this work we present the trisaccharide raffinose as a novel cell culture medium supplement to promote high mannose N-glycans in fed-batch cultures, which is sought after in the development of biosimilars to match the quality profile of the reference medicinal product (RMP) also. Up to six-fold increases of high mannose species were observed with increasing raffinose concentrations in the medium of shaken 96-deepwell plates and shake tubes when culturing two different CHO cell lines in two different media. The findings were confirmed in a pH-, oxygen- and CO2-controlled environment in lab-scale 3.5-L bioreactors. To circumvent detrimental effects on cell growth and productivity at high raffinose concentrations, the media osmolality was adjusted to reach the same value independently of the supplement concentration. Interestingly, raffinose predominantly enhanced mannose 5 glycans, and to a considerably smaller degree, mannose 6. While the underlying mechanism is still not fully understood, minor effects on the nucleotide sugar levels have been observed and transcriptomics analysis revealed that raffinose supplementation altered the expression levels of a number of glycosylation related genes. Among many genes, galactosyltransferase was downregulated and sialyltransferase upregulated. Our results highlight the potential of cell culture medium supplementation to modulate product quality.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 252, 20 June 2017, Pages 32-42
نویسندگان
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