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  • Articles  (16)
  • insect cells  (16)
  • Springer  (16)
  • American Chemical Society
  • 1995-1999  (16)
  • Process Engineering, Biotechnology, Nutrition Technology  (16)
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  • Articles  (16)
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  • 1
    Electronic Resource
    Electronic Resource
    Insect cell physiology (1997)
    Springer
    Cytotechnology 24 (1997), S. 1-9 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; metabolism ; energetics ; amino acids ; fluxes ; oxygen uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/B:CYTO.0000010410.24541.dd
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  • 2
    Electronic Resource
    Electronic Resource
    Efficient membrane targeting of the chick nicotinic acetylcholine receptor α-subunit in stable insect cell lines (1995)
    Springer
    Cytotechnology 19 (1995), S. 37-42 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; membrane-targeting ; nicotinic acetylcholine receptor α-subunit ; Spodoptera frugiperda ; stable expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We have synthesised the α-subunit of the chick nicotinic acetylcholine receptor (nAChR) in stable, continuous insect (Spodoptera frugiperda) cell lines. A cDNA was integrated randomly into the insect cell genome under control of a baculovius immediate early gene promoter. Transformed cells were obtained by co-transfection of the insect cells with pIEK1.nAChRα, encoding the α-subunit cDNA, and pIEK1.neo, encoding the neomycin resistance gene. G-418-resistant clones were selected and expanded into continuous cell lines synthesising the chick nAChR α-subunit. Using fluorescence microscopy and ligand binding studies we were able to demonstrate efficient membrane targeting of the receptor subunit in the insect cell plasma membrane. Stable insect cell lines may thus have significant advantages over transient baculovirus vectors for the synthesis and characterisation of heterologous receptor proteins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749753
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  • 3
    Electronic Resource
    Electronic Resource
    Modelling baculovirus infection of insect cells in culture (1996)
    Springer
    Cytotechnology 20 (1996), S. 209-219 
    Details
    ISSN: 1573-0778
    Keywords: baculovirus ; insect cells ; mathematical model ; population balance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Conclusions Infection of insect cells with baculovirus is a potentially attractive means for producing both viral insecticides and recombinant proteins. The continuation of mathematical modelling studies such as those reviewed in this paper are essential in order to realise the full potential of the system. Through mathematical models it is possible to predict complex behaviours such as those observed when infecting cells at low MOI or when propagating virus in a continuous culture system. A purely empirical analysis of the same phenomena is very difficult if not impossible. The present three models are — despite their complexity and the effort that has gone into developing them — all first generation models. They summarise, to a large extent, our present quantitative understanding of the interaction between baculovirus and insect cells, when looked upon as a black box system. The binding and initial infection processes are still quantitatively poorly understood and further work in this area is much needed. On the longer term, a second generation of models is likely to consider interior processes such as viral DNA and RNA accumulation in much more detail using a structured model of the infection cycle.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350401
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  • 4
    Electronic Resource
    Electronic Resource
    Safety aspects of insect cell culture (1996)
    Springer
    Cytotechnology 20 (1996), S. 299-304 
    Details
    ISSN: 1573-0778
    Keywords: biosafety ; insect cells ; containment ; risk ; virus ; mycoplasma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Conclusions The hazards posed to the environment by the accidental release of baculovirus expression vectors can be put into perspective by the results obtained from experiments in which AcNPV was released deliberately into the field (Bishop et al., 1992). Polyhedrin positive viruses will persist in soil and on leaf surfaces for periods comprising weeks and months. However, polyhedrin negative viruses (similar to those used as expression vectors) do not survive in similar situations. In consequence, accidental release of baculovirus expression vectors poses negligible hazard. The risk of such a release will largely depend on the skill of the operators. This does not take into account the hazard posed by the recombinant product which is being made by the virus-infected insect cell. Synthesis of a mammalian-specific toxin, of course, would require particularly careful manipulation of the virus-infected cell culture. The fact that insect cell lines represent an undefined risk, in terms of carriage of adventitious agents means that their containment should be maintained at a minimum of the European containment level 2. Where the tissue of origin has a high risk of infection with human pathogens or where cells may have been used in a virus culture laboratory then appropriate testing is advisable. Careful risk assessment respecting the scale of work and whole procedures (in addition to individual assessment of agents and reagents) will ensure safe working conditions for laboratory staff. If applied properly safety procedures will also succeed in encouraging clean, efficient and well documented work procedures which are synonymous with the economical use of time and resources and good science.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350409
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  • 5
    Electronic Resource
    Electronic Resource
    Effect of lipids on insect cell growth and expression of recombinant proteins in serum-free medium (1996)
    Springer
    Cytotechnology 22 (1996), S. 211-216 
    Details
    ISSN: 1573-0778
    Keywords: baculovirus ; β-galactosidase ; insect cells ; lipids ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The lipid emulsion components of a serum-free insect cell medium were varied and evaluated for effects on cell growth and recombinant protein expression. The growth of High-FiveTM cells was significantly affected by polyol Pluronic F-68 and Tween-80, but not by lipids. Pluronic was essential for cell growth, while Tween-80 was required to achieve maximum cell densities. A dose response effect was observed for Tween-80 with optimal cell growth at a concentration of 25 mg/l. Cholesterol had a minor effect on cell growth, but was essential for the expression of recombinant proteins. The expression of β-galactosidase (β-gal) was directly affected by cholesterol with optimal expression at a concentration of 5.4 mg/l. Vitamin E, important as an antioxidant to stabilize lipids, did not directly affect recombinant protein expression. Although lipids were not required for cell growth, the presence of lipids were required during the cell growth phase in order to achieve efficient infection with baculovirus. These studies help to define the important components, and range of concentrations, for lipid emulsions which can effectively replace serum in insect cell culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00353941
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  • 6
    Electronic Resource
    Electronic Resource
    Concentrating Autographa californica nuclear polyhedrosis virus and recombinant alkaline phosphatase from insect cells using a temperature-sensitive hydrogel (1997)
    Springer
    Cytotechnology 25 (1997), S. 227-230 
    Details
    ISSN: 1573-0778
    Keywords: Autographa california nuclear polyhedrosis viruses ; insect cells ; recombinant alkaline phosphatase ; separation efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Recombinant alkaline phosphatase expressed in insect cells was concentrated by a factor of one and half times at a separation efficiency of 54.2% using hydrogel ultrafiltration. Enzyme concentration was confirmed by SDS-PAGE as well as by spectrophotometric measurement. Wild and recombinant Autographa californica nuclear polyhedrosis viruses (AcNPV) were concentrated 1.4 and 1.6 times of the feed solution at 48.5 and 60.0% separation efficiency, respectively. Hydrogel ultrafiltration appears to be an attractive alternative for the concentration of AcNPV and recombinant proteins from insect cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007902119501
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  • 7
    Electronic Resource
    Electronic Resource
    Induction of stress proteins in anoxic and hyperthermicSpodoptera frugiperda cells (1995)
    Springer
    Cytotechnology 17 (1995), S. 91-101 
    Details
    ISSN: 1573-0778
    Keywords: anoxia ; hyperthermia ; insect cells ; stress proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract In this study, we compare stress protein induction in anoxic and hyperthermicSpodoptera frugiperda cells. Anoxia transiently induces a cluster of heat shock proteins at 71 and 72 kDa. This is a subset of a larger group of stress proteins induced by heat shock. Several heat shock proteins reported in this study were previously undetected inS. frugiperda. With these additional proteins, the stress response of hyperthermicS. frugiperda closely resembles that ofDrosophila melanogaster. Prior investigations of stress protein induction during oxygen deprivation focused on mammalian cells. In sharp contrast to these cells, anoxicS. frugiperda cells neither induce glucose-regulated proteins nor suppress the heat shock family of 71/72 kDa proteins. These findings provide insight into the virtually unexplored area of stress protein induction in anoxic insect cells. In addition, they help to explain the effects of oxygen deprivation on heterologous protein yield from virally infected insect cells and to develop an oxygenregulated promoter for stably transformed insect cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749396
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  • 8
    Electronic Resource
    Electronic Resource
    Development and characterization of insect cell lines (1996)
    Springer
    Cytotechnology 20 (1996), S. 3-11 
    Details
    ISSN: 1573-0778
    Keywords: cell line establishment ; cryopreservation ; insect cells ; development of ; insect cells ; characterization of ; isoenzymes ; lepidopteran cell cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Conclusions With the wide availability of insect cell culture media, it can generally be considered a routine process to develop new cell lines. Exceptions to this statement do exist, of course. Difficulties may arise when attempting to culture a specific cell type. For example, while there are a few cell lines from insect fat body and at least one from the midgut, it may not be possible to obtain cell lines from these tissues from all insect species due to terminal differentiation and other factors. Also, researchers have desired cell lines from certain species, such as the honey bee, for which no success has been obtained. As in the early days of tissue culture, it is difficult to discern why negative results occur. However, as more is learned about the physiology and nutrition of various insects and tissues, we may get clues which will help solve these questions. The remaining chapters in this book will provide the reader with exciting uses for insect cell culture. As I mentioned earlier, the baculovirus expression vector system has provided a stimulus to the field of insect cell culture not seen previously.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350384
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  • 9
    Electronic Resource
    Electronic Resource
    Insect cell physiology (1996)
    Springer
    Cytotechnology 20 (1996), S. 33-41 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; metabolism ; energetics ; amino acids ; fluxes ; oxygen uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350387
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  • 10
    Electronic Resource
    Electronic Resource
    Post-translational modifications in insect cells (1996)
    Springer
    Cytotechnology 20 (1996), S. 139-144 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; proteins ; post-translational modification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350394
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  • 11
    Electronic Resource
    Electronic Resource
    Shear sensitivity of insect cells (1996)
    Springer
    Cytotechnology 20 (1996), S. 163-171 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; shear sensitivity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Conclusions While insect cells can be easily damaged in bioreactors as a result of hydrodynamic forces, it is also relatively easy to prevent this damage. Of several possible damage mechanisms, the best understood and preventable is the attachment of cells to gas-liquid interfaces and the subjection of these attached cells to the hydro-dynamic forces and/or physical forces associated with these interfaces. For example, cells attached to gas bubbles in a bioreactor can be transported into the foam layer where they are physically removed from the cell suspension, or they can be killed when the gas bubble they are attached to ruptures at the medium-air interface at the top of the bioreactor. The easiest method to prevent this damage is through the use of specific surface active compounds, such as Pluronic F-68 or Methocel E-50 which prevent the cells from attaching to the gas-medium interface.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350397
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  • 12
    Electronic Resource
    Electronic Resource
    Insect cell bioreactors (1996)
    Springer
    Cytotechnology 20 (1996), S. 173-189 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; bioreactors ; parameters ; production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350398
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  • 13
    Electronic Resource
    Electronic Resource
    Scale up aspects of sparged insect-cell bioreactors (1996)
    Springer
    Cytotechnology 20 (1996), S. 221-229 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; bioreactors ; shear ; stirred vessel ; buble column ; airlift ; scale-up
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Conclusion In this chapter we have attempted to evaluate the most important parameters which can be useful for the pur-pose of design and scale up. Insect cells and animal cells in general can be grown well in large vessels. However, none of the theories and parameters discussed in this chapter have been validated on a larger scale than laboratory and small pilot reactors. Selection of the most suitable design and scale-up method there-fore needs in particular studies in larger vessels. The Kolmogorov theory and the killing-volume model are in this respect the most promising approaches for the optimal design of large-scale animal-cell bioreactors.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350402
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  • 14
    Electronic Resource
    Electronic Resource
    Parvovirus diagnostics and vaccine production in insect cells (1996)
    Springer
    Cytotechnology 20 (1996), S. 261-270 
    Details
    ISSN: 1573-0778
    Keywords: insect cells ; parvoviruses ; virion ; immune response ; capsid proteins ; VLPs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00350405
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  • 15
    Electronic Resource
    Electronic Resource
    Enhanced secretory ability for the human factor VIII light chain produced in baculovirus-infected insect cells (1996)
    Springer
    Cytotechnology 21 (1996), S. 165-170 
    Details
    ISSN: 1573-0778
    Keywords: factor VIII ; light chain ; heavy chain ; baculovirus expression system ; insect cells ; enhanced secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The light and truncated heavy chains of human factor VIII, expressed separately in baculovirus-infected insect cells, exhibited different secretory behaviour when compared with each other and with a biologically active fusion molecule of the truncated heavy and light chains. The light chain was very efficiently secreted into culture medium, as judged by high extracellular protein levels and the absence of evidence for light chain retention within cells. Alternatively, proteins containing the heavy chain sequence were poorly secreted and appeared to be sequestered within cells, suggesting that regions within the heavy chain are responsible for the low levels of secreted protein which have generally been observed for recombinant factor VIII.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02215666
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  • 16
    Electronic Resource
    Electronic Resource
    The indirect effects of multiplicity of infection on baculovirus expressed proteins in insect cells: secreted and non-secreted products (1997)
    Springer
    Cytotechnology 24 (1997), S. 73-81 
    Details
    ISSN: 1573-0778
    Keywords: baculovirus ; multiplicity of infection ; time of infection ; time of harvest ; apolipoprotein E ; insect cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The baculovirus expression vector system was employed to produce human apolipoprotein E and β-galactosidase in order to study the effect of multiplicity of infection on secreted and non-secreted recombinant protein production. Prior knowledge of the influence of other cell culture and infection parameters, such as the cell density at time of infection and the time of harvest, allowed determination of the direct and indirect influences of multiplicity of infection on recombinant protein synthesis and degradation in insect cells. Under non-limited, controlled conditions, the direct effect of multiplicity of infection (10−1−10 pfu/cell) on specific recombinant product yields of non-secreted β-galactosidase was found to be insignificant. Instead, the observed increased in accumulated product was directly correlated to the total number of infected cells during the production period and therefore ultimately dependent on an adequate supply of nutrients. Only the timing of recombinant virus and protein production was influenced by, and dependent on the multiplicity of infection. Evidence is presented in this study that indicates the extremely limited predictability of post-infection cell growth at very low multiplicities of infection of less than 0.1 pfu/cell. Due to the inaccuracy of the current virus quantification techniques, combined with the sensitivity of post-infection cell growth at low MOI, the possibility of excessive post-infection cell growth and subsequent nutrient limitation was found to be significantly increased. Finally, as an example, the degree of product stability and cellular and viral protein contamination at low multiplicity of infection is investigated for a secreted recombinant form of human apolipoprotein E. Comparison of human apolipoprotein E production and secretion at multiplicities of infection of 10−4−10 pfu/cell revealed increased product degradation and contamination with intracellular proteins at low multiplicities of infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007962903435
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