Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein

Munzert E, Heidemann R, Büntemeyer H, Lehmann J, Müthing J (1997)
BIOTECHNOLOGY AND BIOENGINEERING 56(4): 441-448.

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Chinese hamster ovary (CHO) cells producing the recombinant glycoprotein human antithrombin III (rhAT III) were batch cultivated in a 20-L bioreactor for 13 days. Neuraminidase activity in cell-free supernatant was monitored during cultivation and free sialic acid was determined by HPLC. Neu5Ac alpha(2-->3)Gal-specific Maackia amurensis and Gal beta(1-->4)GlcNAc-specific Datura stramonium agglutinin were used for determination of sialylated and desialylated rhAT III, respectively. A commercial test kit was used for evaluation of functional rhAT III activity. Supernatant neuraminidase as well as lactate dehydrogenase activity increased significantly during batch growth. The enhanced number of dead cells correlated with increased neuraminidase activity, which seemed to be principally due to cell lysis, resulting in release of cytosolic neuraminidase. Loss of terminally alpha(2-->3) linked sialic acids of the oligosaccharide portions of rhAT III, analyzed in lectin-based Western blot and lectin-adsorbent assays, correlated with a decrease of activity of rhAT III produced throughout long-term batch cultivation. Thus, structural oligosaccharide integrity as well as the functional activity of recombinant glycoprotein depend on the viability and mortality of the bioreactor culture, and batches with a high number of viable cells are required to guarantee production of glycoproteins with maximum biological activity. (C) 1997 John Wiley & Sons, Inc.
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Munzert E, Heidemann R, Büntemeyer H, Lehmann J, Müthing J. Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein. BIOTECHNOLOGY AND BIOENGINEERING. 1997;56(4):441-448.
Munzert, E., Heidemann, R., Büntemeyer, H., Lehmann, J., & Müthing, J. (1997). Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein. BIOTECHNOLOGY AND BIOENGINEERING, 56(4), 441-448.
Munzert, E., Heidemann, R., Büntemeyer, H., Lehmann, J., and Müthing, J. (1997). Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein. BIOTECHNOLOGY AND BIOENGINEERING 56, 441-448.
Munzert, E., et al., 1997. Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein. BIOTECHNOLOGY AND BIOENGINEERING, 56(4), p 441-448.
E. Munzert, et al., “Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein”, BIOTECHNOLOGY AND BIOENGINEERING, vol. 56, 1997, pp. 441-448.
Munzert, E., Heidemann, R., Büntemeyer, H., Lehmann, J., Müthing, J.: Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein. BIOTECHNOLOGY AND BIOENGINEERING. 56, 441-448 (1997).
Munzert, E, Heidemann, R, Büntemeyer, Heino, Lehmann, J, and Müthing, Johannes. “Production of recombinant human antithrombin III on 20-L bioreactor scale: Correlation of supernatant neuraminidase activity, desialylation, and decrease of biological activity of recombinant glycoprotein”. BIOTECHNOLOGY AND BIOENGINEERING 56.4 (1997): 441-448.
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Data provided by Europe PubMed Central.

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PMID: 12783488
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PMID: 11831754
Recombinant glycoprotein product quality in proliferation-controlled BHK-21 cells.
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PMID: 10099468

42 References

Data provided by Europe PubMed Central.

Capillary electrophoresis of carbohydrates.
Oefner PJ, Chiesa C., Glycobiology 4(4), 1994
PMID: 7827401

Olds, Semin. Thromb. Hemostasis 20(), 1994
Fluorometric assay of neuraminidase with a sodium (4-methylumbelliferyl-alpha-D-N-acetylneuraminate) substrate.
Potier M, Mameli L, Belisle M, Dallaire L, Melancon SB., Anal. Biochem. 94(2), 1979
PMID: 464297
Cerebellar granule cells in culture exhibit a ganglioside-sialidase presumably linked to the plasma membrane.
Riboni L, Prinetti A, Bassi R, Tettamanti G., FEBS Lett. 287(1-2), 1991
PMID: 1879535

Schauer, Trends Biochem. Sci. 10(), 1985
Sialic acids as antigenic determinants of complex carbohydrates.
Schauer R., Adv. Exp. Med. Biol. 228(), 1988
PMID: 2459931

Sliwkowski, J. Cell. Biochem. Suppl. 16(), 1992
Extracellular sialidases.
Sweeley CC., Adv. Lipid Res. 26(), 1993
PMID: 8379452
Separation of branched sialylated oligosaccharides using high-pH anion-exchange chromatography with pulsed amperometric detection.
Townsend RR, Hardy MR, Cumming DA, Carver JP, Bendiak B., Anal. Biochem. 182(1), 1989
PMID: 2481411
Sialidase activities of cultured human fibroblasts and the metabolism of GM3 ganglioside.
Usuki S, Lyu SC, Sweeley CC., J. Biol. Chem. 263(14), 1988
PMID: 3129431

Usuki, Ind. J. Biochem. Biophys. 25(), 1988

Warner, Glycobiology 3(), 1993
Characterization of recombinant human antithrombin III synthesized in Chinese hamster ovary cells.
Zettlmeissl G, Conradt HS, Nimtz M, Karges HE., J. Biol. Chem. 264(35), 1989
PMID: 2592368

Zettlmeissl, Bio/Technology 5(), 1987

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