Cell culture media supplemented with raffinose reproducibly enhances high mannose glycan formation
메타 데이터
바이오화학분류
바이오플라스틱
기타
바이오정밀화학
기타
화장품용 기능성소재
기타
의료용 화학소재
식품첨가제
논문
Cell culture media supplemented with raffinose reproducibly enhances high mannose glycan formation
학술지
Journal of biotechnology
저자명
Bruhlmann, D.; Muhr, A.; Parker, R.; Vuillemin, T.; Bucsella, B.; Kalman, F.; Torre, S.; La Neve, F.; Lembo, A.; Haas, T.; Sauer, M.; Souquet, J.; Broly, H.; Hemberger, J.; Jordan, M.
초록
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 CO<SUB>2</SUB>-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.