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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Zeitschriftentitel
BIOTECHNOLOGY AND BIOENGINEERING
Band
56
Ausgabe
4
Seite(n)
441-448
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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. doi:10.1002/(SICI)1097-0290(19971120)56:4<441::AID-BIT9>3.0.CO;2-O
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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PMID: 12783488
Recovery of antithrombin III from milk by expanded bed chromatography.
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PMID: 11831754
Recombinant glycoprotein product quality in proliferation-controlled BHK-21 cells.
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PMID: 10516578
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PMID: 10099468

42 References

Daten bereitgestellt von Europe PubMed Central.

Decreased affinity of recombinant antithrombin for heparin due to increased glycosylation.
Bjork I, Ylinenjarvi K, Olson ST, Hermentin P, Conradt HS, Zettlmeissl G., Biochem. J. 286 ( Pt 3)(), 1992
PMID: 1417738
Optimization of serum-free fermentation processes for antibody production.
Buntemeyer H, Lutkemeyer D, Lehmann J., Cytotechnology 5(1), 1991
PMID: 1367052
Rapid high-performance liquid chromatographic quantification of recombinant human antithrombin III during production and purification.
Buntemeyer H, Tebbe H, Lutkemeyer D, Lehmann J., J. Chromatogr. B, Biomed. Appl. 662(2), 1994
PMID: 7719477

Cantz, Glycoconjugate J. 10(), 1993

Conzelmann, 1987
Fast sequencing of oligosaccharides: the reagent-array analysis method.
Edge CJ, Rademacher TW, Wormald MR, Parekh RB, Butters TD, Wing DR, Dwek RA., Proc. Natl. Acad. Sci. U.S.A. 89(14), 1992
PMID: 1631129

Fan, Effect of glycosylation differences on heparin binding structure. J. Biol. Chem. 268(), 1993

Ferrari, Glycoconjugate J. 12(), 1995
Synthesis of N-acetylneuraminic acid and of CMP-N-acetylneuraminic acid in the rat liver cell.
Ferwerda W, Blok CM, Van Rinsum J., Biochem. J. 216(1), 1983
PMID: 6651781
Survival of recombinant erythropoietin in the circulation: the role of carbohydrates.
Fukuda MN, Sasaki H, Lopez L, Fukuda M., Blood 73(1), 1989
PMID: 2910371
Structural studies on the carbohydrate portion of human antithrombin III.
Franzen LE, Svensson S, Larm O., J. Biol. Chem. 255(11), 1980
PMID: 7372626
Removal of sialic acid from a glycoprotein in CHO cell culture supernatant by action of an extracellular CHO cell sialidase.
Gramer MJ, Goochee CF, Chock VY, Brousseau DT, Sliwkowski MB., Biotechnology (N.Y.) 13(7), 1995
PMID: 9634806
Structural characterization of glycoprotein carbohydrate chains by using diagoxigenin-labeled lectins on blots.
Haselbeck A, Schickaneder E, von der Eltz H, Hosel W., Anal. Biochem. 191(1), 1990
PMID: 2127661
The Super-Spinner: a low cost animal cell culture bioreactor for the CO2 incubator.
Heidemann R, Riese U, Lutkemeyer D, Buntemeyer H, Lehmann J., Cytotechnology 14(1), 1994
PMID: 7765107
Membrane-bound neuraminidases of rat liver. Neuraminidase activity in Golgi apparatus.
Kishore CS, Tulsiani DR, Bhavanandan VP, Carubelli R., J. Biol. Chem. 250(7), 1975
PMID: 235524

Lehmann, 1988
Purification and characterization of cytosolic sialidase from rat liver.
Miyagi T, Tsuiki S., J. Biol. Chem. 260(11), 1985
PMID: 3997846

Munzert, 1995
Capillary electrophoresis of carbohydrates.
Oefner PJ, Chiesa C., Glycobiology 4(4), 1994
PMID: 7827401
Antithrombin: the principal inhibitor of thrombin.
Olds RJ, Lane DA, Mille B, Chowdhury V, Thein SL., Semin. Thromb. Hemost. 20(4), 1994
PMID: 7899868
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
Isolation and properties of a soluble sialidase from the culture fluid of Chinese hamster ovary cells.
Warner TG, Chang J, Ferrari J, Harris R, McNerney T, Bennett G, Burnier J, Sliwkowski MB., Glycobiology 3(5), 1993
PMID: 8286858
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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