The Cellular Substrate: A Very Important Requirement for Baculovirus in vitro Replication

Miltenburger, H.G.; Naser, W.; Harvey, Jack; Huber, Jürg; Huger, Alois M.

doi:10.1515/znc-1984-9-1021

Cited by 30 publications

(9 citation statements)

References 5 publications

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“…The DpN1 cell line was obtained from eggs of a white mutant of Danaus plexippus (Hughes, unpublished) maintained in laboratory culture as described by Hughes et al (12). The cell line was established following the general procedure of Miltenburger et al (13). Specifically, approximately 250 eggs 24-48 h old were surface sterilized by treatment with 2% commercial bleach (0.1% sodium hypochlorite) for 4 min.…”

Section: Methodsmentioning

confidence: 99%

Novel Insect Cell Line Capable of Complex N‐Glycosylation and Sialylation of Recombinant Proteins

Palomares

Joosten

Hughes

et al. 2003

Biotechnology Progress

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Paucimannose or oligomannose structures are usually attached to glycoproteins produced by insect cells, while mammalian glycoproteins usually have complex glycans. The lack of complex glycosylation has limited the use of the insect cell baculovirus expression vector system (BEVS), despite its high productivity and versatility. The availability of cell lines capable of complex glycosylation can overcome such a problem and potentially increase the utility of BEVS. In this work the capability of two novel cell lines, one from Pseudaletia unipuncta (A7S) and one from Danaus plexippus (DpN1), to produce and glycosylate a recombinant protein (secreted human placental alkaline phosphatase, SeAP) was assessed. SeAP produced by Tn5B1-4 cells at a low passage number (<200) was utilized for comparison. The optimal conditions for the production of SeAP by DpN1 cells were defined, and the glycosylation profiles of SeAP produced by the cell lines were quantitatively determined. Both the A7S and the DpN1 cells produced lower concentrations of SeAP than the Tn5B1-4 cells. Less than 5% of the glycans attached to SeAP produced by the Tn5B1-4 cells had complex forms. Glycans attached to SeAP from A7S cells contained 4% hybrid and 8% complex forms. Galactosylated biantennary structures were identified. Glycans attached to SeAP produced by the DpN1 cell line had 6% hybrid and 26% complex forms. Of the complex forms in SeAP from DpN1, 13% were identified as sialylated glycans. The galactosyltransferase activity of the three cell lines was measured and correlated to their ability to produce complex forms. Even though neither novel cell line produced as much recombinant protein as the Tn5B1-4 cells, the glycosylation of SeAP expressed by both cell lines was more complete. These novel cell lines represent interesting alternatives for the production of complex glycosylated proteins utilizing the BEVS.

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Section: Methodsmentioning

confidence: 99%

Novel Insect Cell Line Capable of Complex N‐Glycosylation and Sialylation of Recombinant Proteins

Palomares

Joosten

Hughes

et al. 2003

Biotechnology Progress

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“…The C. pomonella cells susceptible to CpGV were very slow in growth and the production of virus was low. Replication of a GV in early passages of some lepidopteran cell lines has also been reported, but those cell lines all lost susceptibility to GV infection once the cell lines had been further passaged to become stable cell lines (Miltenburger et al ., ; Granados et al ., ). Why lepidopteran cell lines, including the P. xylostella cell lines reported in this study, are commonly non‐permissive to the homologous GV from the insect has yet to be understood.…”

Section: Discussionmentioning

confidence: 99%

Cell lines from diamondback moth exhibiting differential susceptibility to baculovirus infection and expressing midgut genes

Xiang

Wang

et al. 2017

Insect Science

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Six new cell lines were established from embryonic tissues of the diamondback moth, Plutella xylostella (L.). The cell lines showed differential characteristics, including growth in attachment or in suspension, susceptibility to a baculovirus infection and expression of genes involved in the glucosinolate detoxification pathway in P. xylostella larvae. Five of the cell lines grew attached to the culture flask and one cell line grew unattached as a suspension cell line. The cell lines had population doubling times ranging from 18 to 23 h. Among five of the P. xylostella cell lines examined for infection of a nucleopolyhedrovirus from Autographa californica, AcMNPV, four cell lines were highly susceptible to AcMNPV infection, but one was only semi-permissive to AcMNPV infection. The production of two recombinant proteins, a β-galactosidase of bacterial origin and a secreted alkaline phosphatase of eukaryotic origin, in the P. xylostella cell lines was examined in comparison with that in the cell line Sf9 which is commonly used for recombinant protein production. In the P. xylostella cell lines, expression of three important midgut genes involved in the glucosinolate detoxification pathway, including the glucosinolate sulfatase genes GSS1 and GSS2 and the sulfatase modifying factor gene SUMF1, was detected. The P. xylostella cell lines developed in this study could be useful in in vitro research systems for studying insec-virus interactions and complex molecular mechanisms in glucosinolate detoxification and insect-plant interactions.

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“…The established cell line IZD-Cp 0508 was derived from haemocytes and grown in suspension. All cell lines were maintained in Cp cell culture medium (Miltenburger et al, 1984).…”

Section: Methodsmentioning

confidence: 99%

“…As a rearing stock of codling moth was readily available, many new Cp cell lines were established. CmMNPV replicated in 46 of 81 tested cell lines (Miltenburger et al, 1984). Those cell lines giving the highest polyhedron yield over several passages in vitro were used for a morphological, biochemical and biological characterization of CmMNPV and for a comparison of CmMNPV replication in vivo and in vitro.…”

Section: Introductionmentioning

confidence: 99%

Choristoneura murinana Nuclear Polyhedrosis Virus: Comparative Biochemical and Biological Examination of Replication in vivo and in vitro

Naser

Harvey

Huger

et al. 1987

Journal of General Virology

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SUMMARYAn in vitro replication system for the Choristoneura murinana nuclear polyhedrosis virus (CmMNPV) was established and used (i) to characterize this baculovirus biochemically; (ii) to study the cytoplasmic spindle-shaped inclusions (CSIs) associated with CmMNPV replication; and (iii) to compare the cytopathic changes during CmMNPV replication in vivo and in vitro as well as the properties of virions, polyhedra and CSIs from both systems. It was shown that the processes occurring during, and the products of, CmMNPV replication in vitro closely resemble those in vivo, i.e. in larval hosts. Genome analysis by restriction endonucleases, as well as infectivity studies with polyhedra from both sources did not reveal major differences between virus produced in vivo and that produced in vitro. The CSIs were found exclusively in the cytoplasm of infected cells and were shown to consist of a single protein of Mr 50000. Although the biological significance of these spindles, which are produced in large quantities, is not known, they do not seem to be of importance for the infectivity of this baculovirus in vivo.

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The Cellular Substrate: A Very Important Requirement for Baculovirus in vitro Replication

Cited by 30 publications

References 5 publications

Novel Insect Cell Line Capable of Complex N‐Glycosylation and Sialylation of Recombinant Proteins

Novel Insect Cell Line Capable of Complex N‐Glycosylation and Sialylation of Recombinant Proteins

Cell lines from diamondback moth exhibiting differential susceptibility to baculovirus infection and expressing midgut genes

Choristoneura murinana Nuclear Polyhedrosis Virus: Comparative Biochemical and Biological Examination of Replication in vivo and in vitro

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