Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity

Munzert E, Müthing J, Büntemeyer H, Lehmann J (1996)
BIOTECHNOLOGY PROGRESS 12(4): 559-563.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
Download
Es wurden keine Dateien hochgeladen. Nur Publikationsnachweis!
Autor*in
Abstract / Bemerkung
Sialidase activity in cell-free supernatant of batch-cultivated Chinese hamster ovary (CHO) cells producing human recombinant antithrombin III (rhAT III) was monitored during cultivation using 4-methylumbelliferyl substrate and HPLC for free sialic acid determination. Supernatant sialidase as well as lactate dehydrogenase activity increased significantly during batch growth. The enhanced number of dead cells correlated with increasing sialidase activity which seemed to be principally due to cell lysis, resulting in release of cytosolic sialidase. Loss of terminally alpha(2-->3) bound sialic acids of the oligosaccharides of rhAT III was analyzed in lectin-based Western blot and enzyme-linked lectin assays, using Maackia amurensis and Datura stramonium agglutinins for specific determination of Neu5Ac alpha(2-->3)Gal- and Gal beta(1-->4)GlcNAc-terminated glycoproteins, respectively. Results show a remarkable loss of terminal sialic acids of rhAT III along with decrease in CHO cell viability and concomitant increase of dead cells throughout long-term batch cultivation. To avoid this degradation effect, process parameters forcing high viability are essential and harvesting of culture at an early time even at suboptimal recombinant protein concentrations is highly recommended to avoid product desialylation.
Erscheinungsjahr
1996
Zeitschriftentitel
BIOTECHNOLOGY PROGRESS
Band
12
Ausgabe
4
Seite(n)
559-563
ISSN
8756-7938
Page URI
https://pub.uni-bielefeld.de/record/1638682

Zitieren

Munzert E, Müthing J, Büntemeyer H, Lehmann J. Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity. BIOTECHNOLOGY PROGRESS. 1996;12(4):559-563.
Munzert, E., Müthing, J., Büntemeyer, H., & Lehmann, J. (1996). Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity. BIOTECHNOLOGY PROGRESS, 12(4), 559-563. https://doi.org/10.1021/bp9600086
Munzert, E, Müthing, Johannes, Büntemeyer, Heino, and Lehmann, J. 1996. “Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity”. BIOTECHNOLOGY PROGRESS 12 (4): 559-563.
Munzert, E., Müthing, J., Büntemeyer, H., and Lehmann, J. (1996). Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity. BIOTECHNOLOGY PROGRESS 12, 559-563.
Munzert, E., et al., 1996. Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity. BIOTECHNOLOGY PROGRESS, 12(4), p 559-563.
E. Munzert, et al., “Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity”, BIOTECHNOLOGY PROGRESS, vol. 12, 1996, pp. 559-563.
Munzert, E., Müthing, J., Büntemeyer, H., Lehmann, J.: Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity. BIOTECHNOLOGY PROGRESS. 12, 559-563 (1996).
Munzert, E, Müthing, Johannes, Büntemeyer, Heino, and Lehmann, J. “Sialidase activity in culture fluid of Chinese hamster ovary cells during batch culture and its effect on recombinant human antithrombin III integrity”. BIOTECHNOLOGY PROGRESS 12.4 (1996): 559-563.

23 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Impact of cell culture on recombinant monoclonal antibody product heterogeneity.
Liu H, Nowak C, Shao M, Ponniah G, Neill A., Biotechnol Prog 32(5), 2016
PMID: 27452958
Genetic and functional characterization of the NanA sialidase from Clostridium chauvoei.
Vilei EM, Johansson A, Schlatter Y, Redhead K, Frey J., Vet Res 42(), 2011
PMID: 21314964
Effect of production method and gene amplification on the glycosylation pattern of a secreted reporter protein in CHO cells.
Lipscomb ML, Palomares LA, Hernández V, Ramírez OT, Kompala DS., Biotechnol Prog 21(1), 2005
PMID: 15903239
Effects of buffering conditions and culture pH on production rates and glycosylation of clinical phase I anti-melanoma mouse IgG3 monoclonal antibody R24.
Müthing J, Kemminer SE, Conradt HS, Sagi D, Nimtz M, Kärst U, Peter-Katalinić J., Biotechnol Bioeng 83(3), 2003
PMID: 12783488
Modification of a recombinant GPI-anchored metalloproteinase for secretion alters the protein glycosylation.
Morrison CJ, Easton RL, Morris HR, McMaster WR, Piret JM, Dell A., Biotechnol Bioeng 68(4), 2000
PMID: 10745209
Na-butyrate increases the production and alpha2,6-sialylation of recombinant interferon-gamma expressed by alpha2,6- sialyltransferase engineered CHO cells.
Lamotte D, Buckberry L, Monaco L, Soria M, Jenkins N, Engasser JM, Marc A., Cytotechnology 29(1), 1999
PMID: 19003337
A novel autoregulated proliferation-controlled production process using recombinant CHO cells.
Mazur X, Eppenberger HM, Bailey JE, Fussenegger M., Biotechnol Bioeng 65(2), 1999
PMID: 10458734
Bicarbonate concentration and osmolality are key determinants in the inhibition of CHO cell polysialylation under elevated pCO(2) or pH.
Zanghi JA, Schmelzer AE, Mendoza TP, Knop RH, Miller WM., Biotechnol Bioeng 65(2), 1999
PMID: 10458739
Export

Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Web of Science

Dieser Datensatz im Web of Science®
Quellen

PMID: 8987482
PubMed | Europe PMC

Suchen in

Google Scholar