Baculovirus expression: old dog, new tricks

Berger, Imre; Poterszman, Arnaud

doi:10.1080/21655979.2015.1104433

Cited by 16 publications

(8 citation statements)

References 50 publications

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“…However, selection of the recombinant baculovirus that contains the gene of interest by plaque purification usually takes about 1 week . To alleviate plaque selection and purification recombinant baculovirus are commonly generated using a progenitor baculoviral genome in form of a bacterial artificial chromosome (BAC) and Tn7 transposition in E. coli cells …”

Section: Insect Cell‐baculovirus Platform For Recombinant Vaccine Manmentioning

confidence: 99%

Emerging Technologies for Low‐Cost, Rapid Vaccine Manufacture

Kis

Shattock

Shah

et al. 2018

Biotechnology Journal

117

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To stop the spread of future epidemics and meet infant vaccination demands in low‐ and middle‐income countries, flexible, rapid and low‐cost vaccine development and manufacturing technologies are required. Vaccine development platform technologies that can produce a wide range of vaccines are emerging, including: a) humanized, high‐yield yeast recombinant protein vaccines; b) insect cell‐baculovirus ADDomer vaccines; c) Generalized Modules for Membrane Antigens (GMMA) vaccines; d) RNA vaccines. Herein, existing and future platforms are assessed in terms of addressing challenges of scale, cost, and responsiveness. To assess the risk and feasibility of the four emerging platforms, the following six metrics are applied: 1) technology readiness; 2) technological complexity; 3) ease of scale‐up; 4) flexibility for the manufacturing of a wide range of vaccines; 5) thermostability of the vaccine product at tropical ambient temperatures; and 6) speed of response from threat identification to vaccine deployment. The assessment indicated that technologies in the order of increasing feasibility and decreasing risk are the yeast platform, ADDomer platform, followed by RNA and GMMA platforms. The comparative strengths and weaknesses of each technology are discussed in detail, illustrating the associated development and manufacturing needs and priorities.

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Section: Insect Cell‐baculovirus Platform For Recombinant Vaccine Manmentioning

confidence: 99%

Emerging Technologies for Low‐Cost, Rapid Vaccine Manufacture

Kis

Shattock

Shah

et al. 2018

Biotechnology Journal

117

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“…The CuV and TuV VP2 genes were cloned into the pFastBac1 plasmid to generate recombinant baculoviruses that express virus-like particles (VLPs) using the Bac-to-Bac system according to the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA) [23]. Sf9 insect cells, maintained in SFM Sf9-900 II medium (ThermoFisher, Waltham, MA, USA) supplemented with 1% antibiotic-antimycotic (ThermoFisher) at 28 • C, were infected with the recombinant baculoviruses at a multiplicity of infectivity (MOI) of 5 and harvested 72 h post infection by centrifugation at 2000 rpm for 20 min at 4 • C. The pellets were re-suspended in lysis buffer (25 mM Tris-HCl pH 8.0, 100 mM NaCl, 0.2% Triton X-100, 2 mM MgCl 2 ) (TNTM buffer) and frozen and stored at −20 • C until purification.…”

Section: Production and Purification Of Cuv And Tuv Virus-like Particlesmentioning

confidence: 99%

Structural Characterization of Cuta- and Tusavirus: Insight into Protoparvoviruses Capsid Morphology

Mietzsch

McKenna

Väisänen

et al. 2020

Viruses

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Several members of the Protoparvovirus genus, capable of infecting humans, have been recently discovered, including cutavirus (CuV) and tusavirus (TuV). To begin the characterization of these viruses, we have used cryo-electron microscopy and image reconstruction to determine their capsid structures to ~2.9 Å resolution, and glycan array and cell-based assays to identify glycans utilized for cellular entry. Structural comparisons show that the CuV and TuV capsids share common features with other parvoviruses, including an eight-stranded anti-parallel β-barrel, depressions at the icosahedral 2-fold and surrounding the 5-fold axes, and a channel at the 5-fold axes. However, the viruses exhibit significant topological differences in their viral protein surface loops. These result in three separated 3-fold protrusions, similar to the bufaviruses also infecting humans, suggesting a host-driven structure evolution. The surface loops contain residues involved in receptor binding, cellular trafficking, and antigenic reactivity in other parvoviruses. In addition, terminal sialic acid was identified as the glycan potentially utilized by both CuV and TuV for cellular entry, with TuV showing additional recognition of poly-sialic acid and sialylated Lewis X (sLeXLeXLeX) motifs reported to be upregulated in neurotropic and cancer cells, respectively. These structures provide a platform for annotating the cellular interactions of these human pathogens.

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“…to modify, activate or inactivate a particular protein specimen produced from the Tn7 site on the same baculoviral genome [6,15]. This additional site proved to be highly useful for generating customized MultiBac baculoviral genomes for specific applications (Figure 1) [10,[16][17][18][19][20][21][22]. All along, our ambition was to streamline the handling of the entire system, including composite baculovirus generation, tissue culture, virus amplification and protein complex production, to render the technique accessible to a wide range of users including non-specialists who would only occasionally use this tool.…”

Section: Main Text Multibac Origins: the Nuts And Boltsmentioning

confidence: 99%

The MultiBac system: a perspective

Berger

Tölzer

Gupta

2019

Emerging Topics in Life Sciences

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Baculovirus expression is a time-tested technique to produce proteins in insect cells, in high quality and quantity for a range of applications. MultiBac is a baculovirus expression system we developed originally for producing multiprotein complexes comprising many subunits, for structural and mechanistic studies. First introduced in 2004, MultiBac is now in use in many laboratories worldwide, accelerating research programmes in academia and industry. We have continuously optimized our MultiBac system, providing customized reagents and standard operating protocols to facilitate its use also by non-specialists. More recently, we have generated MultiBac genomes tailored for specific purposes, for example, to produce humanized glycoproteins, high-value pharmaceutical targets including kinases, viral polymerases, and virus-like particles (VLPs) as promising vaccine candidates. By altering the host tropism of the baculovirion, we created MultiBacMam, a heterologous DNA delivery toolkit to target mammalian cells, tissues and organisms. Introducing CRISPR/Cas modalities, we set the stage for large-scale genomic engineering applications utilizing this high-capacity DNA delivery tool. Exploiting synthetic biology approaches and bottom-up design, we engage in optimizing the properties of our baculoviral genome, also to improve manufacturing at scale. Here we provide a perspective of our MultiBac system and its developments, past, present and future.

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Baculovirus expression: old dog, new tricks

Cited by 16 publications

References 50 publications

Emerging Technologies for Low‐Cost, Rapid Vaccine Manufacture

Emerging Technologies for Low‐Cost, Rapid Vaccine Manufacture

Structural Characterization of Cuta- and Tusavirus: Insight into Protoparvoviruses Capsid Morphology

The MultiBac system: a perspective

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