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  • 1
    Electronic Resource
    Electronic Resource
    Glycosylation patterns of the human colon cancer cell line HT-29 detected byHelix pomatia agglutinin and other lectins in culture, in primary tumours and in metastases in SCID mice (1994)
    Springer
    Clinical & experimental metastasis 12 (1994), S. 398-404 
    Details
    ISSN: 1573-7276
    Keywords: colon cancer ; glycosylation ; Helix pomatia agglutinin ; metastasis ; SCID mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Human colonic cancer cells (HT-29, 10 7 cells/dose) were injected subcutaneously between the scapulae of 19 severe combined immunodeficient (SCID) mice. After 19 days, large tumours had developed in 18 out of the 19 animals and the mice were then killed. Metastases were detected in the lungs of 16 animals but not in other organs investigated. Surgical removal of the primary tumour in another group of five animals led to a prolonged survival and further growth of metastases in the lungs. HT-29 injection into the tail vein (n=5)resulted in colonization of the lungs. The tumours that developed in the animals were signet cell carcinomas; these forms are not seen in HT-29 cells in culture. Glycoconjugate expression of the tumours was assessed using several lectins. In many cases the results indicated a stability of lectin-binding patterns from cell culture conditions to implantation into the SCID mice. This was true for the lectin Helix pomatia agglutinin (HPA), the binding of which is associated with a high metastatic potential in some human tumours, including colon cancer. All the primary tumours and metastases were HPA positive. This xenograft tumour model seems to be a clinically relevant system for the study of glycoconjugate expression in human colon cancer cells and their metastases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01755883
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  • 2
    Electronic Resource
    Electronic Resource
    Markers of prognosis in breast cancer - the relationship between binding of the lectin HPA and histological grade, SPF, and ploidy (1993)
    Springer
    Breast cancer research and treatment 25 (1993), S. 247-256 
    Details
    ISSN: 1573-7217
    Keywords: breast cancer ; glycosylation ; HPA ; Helix pomatia lectin ; prognosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Abnormal cellular glycosylation as demonstrated by the binding of a lectin fromHelix pomatia (HPA) to paraffin-embedded sections has been shown in several studies to be associated with aggressive biological behaviour and poor long-term patient prognosis in breast cancer. This study aims to address the possibility that expression of the HPA binding ligand may be of prognostic significance through an association with increased cellular proliferation (as measured by S-phase fraction and histological grade), anaplasia (reflected in histological grade), or ploidy (DNA index). In a 24 year retrospective study, paraffin-embedded sections of 366 primary breast cancers were stained for binding of HPA. All tumours were assessed for histological grade. Flow cytometry was performed on all cases for which sufficient tumour tissue was available (358/366 cases) and S-phase fraction (SPF) and ploidy calculated. Data regarding patient age at diagnosis, nodal status, and tumour size were also recorded. Life table analyses revealed survival advantage for HPA ‘non stainers’ in comparison to ‘stainers’ (p〈 0.001); for patients with tumours of low grade vs. high grade (p〈0.001); and for those with tumours of low SPF vs. high SPF (p〈0.001). No survival advantage was shown for those with diploid vs. aneuploid tumours (p= 0.17). No association was apparent between HPA binding and grade, SPF, or ploidy (Chi squared values not significant). This was confirmed by multivariate analysis in which nodal status, tumour size, and SPF were independently predictive of survival. There was no confounding effect of grade, SPF, or ploidy upon the correlation between survival and HPA binding. HPA was, however, not independently predictive owing to its strong association with nodal status. The results of this study suggest that the prognostic significance of altered glycosylation, as detected by HPA binding, is unlikely to be through an association with proliferative rate, degree of anaplasia, or cellular ploidy, but may rather be through a direct association with the presence of nodal metastases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00689839
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  • 3
    Electronic Resource
    Electronic Resource
    The effect of the dilution rate on CHO cell physiology and recombinant interferon-γ production in glucose-limited chemostat culture (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 42 (1993), S. 1077-1085 
    Details
    ISSN: 0006-3592
    Keywords: chemostat ; glucose limitation ; glycosylation ; CHO cells ; interferon-γ ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The physiology of a recombinant Chinese hamster ovary cell line in glucose-limited chemostat culture was studied over a range of dilution rates (D = 0.008 to 0.20 h-1). The specific growth rate (μ) deviated from D at low dilution rates due to an increased specific death rate. Extrapolation of these data suggested a minimum specific growth rate of 0.011 h-1 (μmax = 0.025 h-1) The metabolism at each steady state was characterized by determining the metabolic quotients for glucose, lactate, ammonia, amino acids, and interferon-γ (IFN-γ). The specific rate of glucose uptake increased linearly with μ, and the saturation constant for glucose (Ks) was calculated to be 59.6 μM. There was a linear increase in the rate of lactate production with a higher yield of lactate from glucose at high growth rates. The decline in the rate of production of lactate, alanine, and serine at low growth rate was consistent with the limitation of the glycolytic pathway by glucose. The specific rate of IFN-γ production increased with μ in a manner indicative of a growth-related product. Despite changes in the IFN-γ production rate and cell physiology, the pattern of IFN-γ glycosylation was similar at all except the lowest growth rates where there was increased production of nonglycosylated IFN-γ. © 1993 John Wiley & Sons, Inc.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260420909
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  • 4
    Electronic Resource
    Electronic Resource
    N-glycans of recombinant human interferon-γ change during batch culture of chinese hamster ovary cells (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 48 (1995), S. 639-648 
    Details
    ISSN: 0006-3592
    Keywords: interferon ; glycosylation ; CHO cells ; microheterogeneity ; mass spectrometry ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A recombinant Chinese hamster ovary (CHO) cell line making human interfron-γ (IFN-γ) was grown in 12-L stirred tank fermentors in three batch fermentations under conditions of constant temperature, pH, and dissolved oxygen tension. In addition to cell growth, metabolite, and productivity data, a detailed analysis of the carbohydrate structures attached to each glycosylation site of IFN-γ was achieved using matrix-assisted laser desorption mass spectrometry (MALDI-MS) in combination with exoglycosidase array sequencing. Complex biantennary oligosaccharides (particularly Gal2GlcNAc4Man3 which was core ℵl-6 fucosylated at Asn25 but not at Asng97) were most prevalent at both glycosylation sites. However, considerable microheterogeneity arising from the presence of triantennary and truncated glycan structures was also observed. The proportion of the dominant core glycan structure (Gal2GlcNAc4Man3 ± Fuc1) decreased by 15-26% during batch culture, with increases in the proportion of oligomannose and truncated glycans over the same time period. Prolonged culture resulting from an extended lag phase led to further accumulation of oligomannose and truncated structures, reaching up to 52% of total glycans attached to Asng97 by 240 h of culture. The implications of these glycosylation changes for optimizing the time for harvesting cell cultures, and for the clearance of recombinant therapeutic products in vivo are discussed. © 1995 John Wiley & Sons, Inc.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260480612
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  • 5
    Electronic Resource
    Electronic Resource
    Glucose-limited chemostat culture of chinese hamster ovary cells producing recombinant human interferon-γ (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 39 (1992), S. 327-335 
    Details
    ISSN: 0006-3592
    Keywords: Chinese hamster ovary ; interferon-γ ; chemostat culture ; glycosylation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A Chinese hamster ovary (CHO) cell line expressing recombinant human interferon-γ (IFN-γ) was grown under glucose limitation in a chemostate at a constant dilution rate of 0.015 h-1 with glucose feed concentrations of 2.75 mM and 4.25 mM. The changes in cell concentration that accompanied changes in the glucose feed concentration indicated that the cells were glucose-limited. The cell yield on glucose remained constant, but there was a decline in residual glucose concentration and a reduced lactate yield from glucose in the latter stages of the culture. The consumption rates for many of the essential amino acids were increased later in the culture. The volumetric rate of interferon-γ production was maintained throughout the course of this culture, indicating that IFN-γ expression was stable under these conditions. However, the specific rate of IFN-γ production was significantly lower at the higher glucose feed concentration. Under glucose limitation, the proportion of fully glycosylated IFN-γ produced by these cells was less than that produced in the early stages of batch cultures. The proportion of fully glycosylated IFN-γ increased during transient periods of glucose excess, suggesting that the culture environment influences the glycosylation of IFN-γ.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260390311
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