Glycosidase Activities in Chinese Hamster Ovary Cell Lysate and Cell Culture Supernatant

Gramer, Michael J.; Goochee, Charles F.

doi:10.1021/bp00022a003

Cited by 137 publications

(111 citation statements)

References 45 publications

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“…This effect explains the decline of neuraminidase activity during late batch cultivation. Thus, our findings concerning soluble neuraminidase activity in CHO cell culture supernatant are in good agreement with data from other groups who analyzed glycosidase activities in the culture fluid and in lysates of CHO cells during batch and continuous culture in previous studies (Gramer and Goochee, 1993;Gramer et al, 1995;Warner et al, 1993). Compared to data from Gramer et al (1995), the batch cultivation of this report was shorter (6 days) with rising LDH activity from day 3 after inoculation.…”

Section: Neuraminidase Activity In Bioreactor Supernatant Of Rhat IIIsupporting

confidence: 92%

“…Moreover, neuraminidase activity has been detected in the cell-free culture fluid of mammalian cells and cell lines (Gramer and Goochee, 1993;Gramer et al, 1995;Sweeley, 1993). A soluble neuraminidase has been purified from the supernatant of Chinese hamster ovary (CHO) cells, and this enzyme has recently been cloned (Ferrari et al, 1994;Warner et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

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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

Heidemann²,

Lehmann³

et al. 1997

Biotechnol. Bioeng.

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Section: Neuraminidase Activity In Bioreactor Supernatant Of Rhat IIIsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

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

Heidemann²,

Lehmann³

et al. 1997

Biotechnol. Bioeng.

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“…Desialylation in presence of serum could also be due to an enzymatic degradation of protein glycans by serum enzymes or by cytoplasmic enzymes released during cell lysis (Gramer and Goochee 1993). In the case of sialidase activities present in serum preparations (Gawlitzek et al 1995), a protein desialylation should have appeared from the early beginning of the culture.…”

Section: Discussionmentioning

confidence: 99%

Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells

et al. 2006

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The necessity to perform serum-free cultures to produce recombinant glycoproteins generally requires an adaptation procedure of the cell line to new environmental conditions, which may therefore induce quantitative and qualitative effects on the product, particularly on its glycosylation. In previous studies, desialylation of EPO produced by CHO cells was shown to be dependent on the presence of serum in the medium. In this paper, to discriminate between the effects of the adaptation procedure to serum-free medium and the effects of the absence of serum on EPO production and glycosylation, adapted and nonadapted CHO cells were grown in serum-free and serum-containing media. The main kinetics of CHO cells were determined over batch processes as well as the glycosylation patterns of produced EPO by HPCE-LIF. A reversible decrease in EPO production was observed when cells were adapted to SFX-CHO TM medium, as the same cells partially recovered their production capacity when cultivated in serum-containing medium or in the enriched SFM TM serum-free medium. More interestingly, EPO desialylation that was not observed in both serum-free media was restored if the serum-independent cells were recultured in presence of serum. In the same way, while the serum-independent cells did not release a sialidase activity in both serum-free media, a significant activity was recovered when serum was added. In fact, the cell adaptation process to serum-free conditions did not specifically affect the sialidase release and the cellular mechanism of protein desialylation, which appeared to be mainly related to the presence of serum for both adapted and non-adapted cells.

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“…All of the purification steps were carried out at 4°C. ␣-L-Fucosidase activity was carried out as described (41) …”

Section: Methodsmentioning

confidence: 99%

Role for α- l -fucosidase in the control of Helicobacter pylori -infected gastric cancer cells

Liu

Huang

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Infecting about one-half of the global human population, Helicobacter pylori is well established as the primary cause of gastritis, duodenal ulcer, and gastric cancer. Currently there is no clear information regarding if and how host cells interact with H. pylori, and if such interactions are dependent on the type of gastric disease. Using fluorescently labeled fucose-containing glycoconjugates, we provide evidence observing both the uptake of L-fucose from gastric cancer cells to H. pylori and that human ␣-L-fucosidase 2 (FUCA2) is secreted only under coculture conditions (i.e., host cells infected with H. pylori). Upon depletion of FUCA2 by RNA interference and detection of translocated CagA (a virulence factor of H. pylori) in host cells, FUCA2 was found to be essential for H. pylori adhesion, in particular to the gastric cancer-and duodenal ulcer-specific strains. Additionally FUCA2 was shown to significantly enhance the expression of Lewis x antigen in H. pylori, which is critical for bacterial cell adhesion in the pathogenesis and defense strategy to escape host surveillance. These findings not only demonstrate an important connection between FUCA2 and the adhesion, growth, and pathogenicity of H. pylori, but also support the idea that FUCA2 is a potential target for clinical diagnosis and therapeutic intervention of H. pylori-related diseases.

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Glycosidase Activities in Chinese Hamster Ovary Cell Lysate and Cell Culture Supernatant

Cited by 137 publications

References 45 publications

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

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

Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells

Role for α- l -fucosidase in the control of Helicobacter pylori -infected gastric cancer cells

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