Large-Scale Propagation of Insect Cells

2018 ◽  
pp. 597-618 ◽  
Author(s):  
James L. Vaughn ◽  
Stefan A. Weiss
Keyword(s):  
Large Scale ◽  
Insect Cells

Industrial Large Scale of Suspension Culture of Insect Cells

2014 ◽  
pp. 348-389 ◽  
Author(s):  
António Roldão ◽  
Manon Cox ◽  
Paula Alves ◽  
Manuel Carrondo ◽  
Tiago Vicente
Keyword(s):  
Suspension Culture ◽  
Large Scale ◽  
Insect Cells

Enhanced Expression, Secretion, and Large-Scale Purification of Recombinant HIV-1 gp120 in Insect Cells Using the Baculovirus egt and p67 Signal Peptides

1993 ◽  
Vol 4 (5) ◽  
pp. 349-357 ◽  
Author(s):  
C.I. Murphy ◽  
J.R. Mcintire ◽  
D.V. Davis ◽  
H. Hodgdon ◽  
J.R. Seals ◽  
...  
Keyword(s):  
Large Scale ◽  
Insect Cells ◽  
Signal Peptides ◽  
Enhanced Expression ◽  
Hiv 1

scholarly journals Heterelogous Expression of Plant Genes

2009 ◽  
Vol 2009 ◽  
pp. 1-16 ◽  
Author(s):  
Filiz Yesilirmak ◽  
Zehra Sayers
Keyword(s):  
Large Scale ◽  
Protein Production ◽  
Insect Cells ◽  
Protein Characterization ◽  
Model Organisms ◽  
Plant Proteins ◽  
Production Methods ◽  
Functional Studies ◽  
Recombinant Plant ◽  
Plant Genes

Heterologous expression allows the production of plant proteins in an organism which is simpler than the natural source. This technology is widely used for large-scale purification of plant proteins from microorganisms for biochemical and biophysical analyses. Additionally expression in well-defined model organisms provides insights into the functions of proteins in complex pathways. The present review gives an overview of recombinant plant protein production methods using bacteria, yeast, insect cells, and Xenopus laevis oocytes and discusses the advantages of each system for functional studies and protein characterization.

Structural implications for the transformation of the Bacillus thuringiensis δ-endotoxins from water-soluble to membrane-inserted forms

2001 ◽  
Vol 29 (4) ◽  
pp. 571-577 ◽  
Author(s):  
J. Li ◽  
D. J. Derbyshire ◽  
B. Promdonkoy ◽  
D. J. Ellar
Keyword(s):  
Bacillus Thuringiensis ◽  
Conformational Change ◽  
Conformational Changes ◽  
Large Scale ◽  
Insect Cells ◽  
Water Soluble ◽  
Single Amino Acid ◽  
The Core ◽  
High Affinity Site ◽  
Temperature Factors

Crystal structures combined with biochemical data show that the δ-endotoxins from Bacillus thuringiensis are structurally poised towards large-scale, irreversible conformational changes that transform them from the soluble protein bound at the cell surface into a membrane-embedded form causing lysis of susceptible insect cells. Cry δ-endotoxins are made of a helix bundle, a β-prism and a β-sandwich. The conformational change involves an umbrella-like opening between the helix-4,5-hairpin and the remaining helices, and between the helical domain and the two sheet domains. Comparison of Cry1Ac structures with and without the bound receptor ligand GalNAc associates occupation of the high-affinity site on the β-sandwich with an increase of temperature factors in the helical, pore-forming domain, which may indicate how receptor binding could trigger the required major conformational change. The structure of Cyt δ-endotoxins indicates that the surface helix hairpins must peel away to expose the β-strands for membrane attack. Single amino acid substitutions in hinge residues or the core can restore activity following an inhibitory mutation.

scholarly journals Spontaneous excision of BAC vector sequences from bacmid-derived baculovirus expression vectors upon passage in insect cells

2003 ◽  
Vol 84 (10) ◽  
pp. 2669-2678 ◽  
Author(s):  
Gorben P. Pijlman ◽  
Jessica E. van Schijndel ◽  
Just M. Vlak
Keyword(s):  
Protein Expression ◽  
Viral Genome ◽  
Large Scale ◽  
Heterologous Protein ◽  
Insect Cells ◽  
Expression Cassette ◽  
Expression Vectors ◽  
Green Fluorescent Protein Gene ◽  
Heterologous Protein Expression ◽  
Scale Production

Repeated baculovirus infections in cultured insect cells lead to the generation of defective interfering viruses (DIs), which accumulate at the expense of the intact helper virus and compromise heterologous protein expression. In particular, Autographa californica multicapsid nucleopolyhedovirus (AcMNPV) DIs are enriched in an origin of viral DNA replication (ori) not associated with the homologous regions (hrs). This non-hr ori is located within the coding sequence of the non-essential p94 gene. We investigated the effect of a deletion of the AcMNPV non-hr ori on the heterologous protein expression levels following serial passage in Sf21 insect cells. Using homologous ET recombination in E. coli, deletions within the p94 gene were made in a bacterial artificial chromosome (BAC) containing the entire AcMNPV genome (bacmid). All bacmids were equipped with an expression cassette containing the green fluorescent protein gene and a gene encoding the classical swine fever virus E2 glycoprotein (CSFV-E2). For the parental (intact) bacmid only, a strong accumulation of DIs with reiterated non-hr oris was observed. This was not observed for the mutants, indicating that removal of the non-hr ori enhanced the genetic stability of the viral genome upon passaging. However, for all passaged viruses it was found that the entire BAC vector including the expression cassette was spontaneously deleted from the viral genome, leading to a rapid decrease in GFP and CSFV-E2 production. The rationale for the (intrinsic) genetic instability of the BAC vector in insect cells and the implications with respect to large-scale production of proteins with bacmid-derived baculoviruses are discussed.

scholarly journals Over-expression and characterization of active recombinant rat liver carnitine palmitoyltransferase II using baculovirus

1995 ◽  
Vol 309 (2) ◽  
pp. 689-693 ◽  
Author(s):  
T M Johnson ◽  
W R Mann ◽  
C J Dragland ◽  
R C Anderson ◽  
G M Nemecek ◽  
...  
Keyword(s):  
Rat Liver ◽  
Large Scale ◽  
Insect Cells ◽  
Specific Activity ◽  
Recombinant Baculovirus ◽  
Cell System ◽  
Carnitine Palmitoyltransferase ◽  
Sds Page ◽  
Infected Insect ◽  
Carnitine Palmitoyltransferase Ii

The cDNA encoding rat liver carnitine palmitoyltransferase II (CPT-II) was heterologously expressed using a recombinant baculovirus/insect cell system. Unlike Escherichia coli, the baculovirus-infected insect cells expressed mostly soluble active recombinant CPT-II (rCPT-II). CPT activity from crude lysates of recombinant baculovirus-infected insect cells was maximal between 50 and 72 h post-infection, with a peak specific activity of 100-200 times that found in the mock- or wild-type-infected control lysates. Milligram quantities (up to 1.8 mg/l of culture) of active rCPT-II were chromatographically purified from large-scale cultures of insect cells infected with the recombinant baculovirus. The rCPT-II was found to be: (1) similar in size to the native rat liver enzyme (approximately 70 kDa) as judged by SDS/PAGE; (2) immunoreactive with a polyclonal serum raised against rat liver CPT-II; and (3) not glycosylated. Kinetic analysis of soluble rCPT-II revealed Km values for carnitine and palmitoyl-CoA of 950 +/- 27 microM and 34 +/- 5.6 microM respectively.

scholarly journals Large-scale recoding of an arbovirus genome to rebalance its insect versus mammalian preference

2015 ◽  
Vol 112 (15) ◽  
pp. 4749-4754 ◽  
Author(s):  
Sam H. Shen ◽  
Charles B. Stauft ◽  
Oleksandr Gorbatsevych ◽  
Yutong Song ◽  
Charles B. Ward ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Large Scale ◽  
Insect Cells ◽  
Animal Kingdom ◽  
Specific Expression ◽  
Live Attenuated Vaccine ◽  
Newborn Mice ◽  
Protective Antibodies ◽  
Attenuated Viruses ◽  
New Generation

The protein synthesis machineries of two distinct phyla of the Animal kingdom, insects of Arthropoda and mammals of Chordata, have different preferences for how to best encode proteins. Nevertheless, arboviruses (arthropod-borne viruses) are capable of infecting both mammals and insects just like arboviruses that use insect vectors to infect plants. These organisms have evolved carefully balanced genomes that can efficiently use the translational machineries of different phyla, even if the phyla belong to different kingdoms. Using dengue virus as an example, we have undone the genome encoding balance and specifically shifted the encoding preference away from mammals. These mammalian-attenuated viruses grow to high titers in insect cells but low titers in mammalian cells, have dramatically increased LD50s in newborn mice, and induce high levels of protective antibodies. Recoded arboviruses with a bias toward phylum-specific expression could form the basis of a new generation of live attenuated vaccine candidates.

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