Insect larvae biofactories as a platform for influenza vaccine production

Gómez-Casado, Eduardo; Gómez-Sebastián, Silvia; Núñez, María; Lasa, Rodrigo; Martínez-Pulgarín, Susana; Escribano, José M.

doi:10.1016/j.pep.2011.03.007

Cited by 28 publications

(17 citation statements)

References 53 publications

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“…A variety of transfer vectors encoding resident fusion proteins reported to improve protein expression are now available for the construction of recombinant baculoviruses. These include maltose binding protein [10], glutathione S transferase [11], SUMO [12] and KDEL retention signal [13]. The deletion of other non-essential virus genes encoding for proteins such as p26, p10 and p74 also offer advantages for productivity [14].…”

Section: Introductionmentioning

confidence: 99%

Significant Productivity Improvement of the Baculovirus Expression Vector System by Engineering a Novel Expression Cassette

Gómez-Sebastián¹,

López-Vidal²,

Escribano

2014

PLoS ONE

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Here we describe the development of a baculovirus vector expression cassette containing rearranged baculovirus-derived genetic regulatory elements. This newly designed expression cassette conferred significant production improvements to the baculovirus expression vector system (BEVS), including prolonged cell integrity after infection, improved protein integrity, and around 4-fold increase in recombinant protein production yields in insect cells with respect to a standard baculovirus vector. The expression cassette consisted of a cDNA encoding for the baculovirus transactivation factors IE1 and IE0, expressed under the control of the polyhedrin promoter, and a homologous repeated transcription enhancer sequence operatively cis-linked to the p10 promoter or to chimeric promoters containing p10. The prolonged cell integrity observed in cells infected by baculoviruses harbouring the novel expression cassette reduced the characteristic proteolysis and aberrant forms frequently found in baculovirus-derived recombinant proteins. The new expression cassette developed here has the potential to significantly improve the productivity of the BEVS.

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Section: Introductionmentioning

confidence: 99%

Significant Productivity Improvement of the Baculovirus Expression Vector System by Engineering a Novel Expression Cassette

Gómez-Sebastián¹,

López-Vidal²,

Escribano

2014

PLoS ONE

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“…These results are in agreement with a previous report showing that hemagglutinin was expressed at ca. 65 kDa in Trichoplusia ni larvae when using a baculovirus system (Gomez-Casado et al, 2011), and also indicate that rHA expressed 3) were immobilized on microplates, and the protein solution containing hemolymph and fractions obtained by affinity and gel filtration chromatography were added to wells of a microplate. After washing, rabbit polyclonal antibody to influenza A virus H5N1 (avian flu) HA was added as a primary antibody, and a goat anti-Rabbit IgG-HRP was added as a secondary antibody.…”

Section: Discussionmentioning

confidence: 99%

Expression and purification of bioactive hemagglutinin protein of highly pathogenic avian influenza A (H5N1) in silkworm larvae

Dong¹,

Harada

Yoshida

et al. 2013

Journal of Virological Methods

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The hemagglutinin (HA) of avian influenza viruses plays a very important role in the infection of host cells. In this study, the HA gene of the highly pathogenic avian influenza H5N1 virus was cloned and expressed in silkworm larvae. The expressed recombinant HA (rHA) was purified using fetuin-agarose chromatography and Superdex 200 10/300 GL gel filtration chromatography, and the identity of purified rHA was confirmed by SDS-PAGE and Western blot. Approximately 500 μg of purified rHA was obtained from a total of 30 silkworm larvae, suggesting the high efficiency of the silkworm expression system. The purified rHA bound to a rabbit polyclonal antibody against influenza A virus H5N1 (avian flu) HA, suggesting its antigenicity and potential application in vaccine development. Gel filtration chromatography showed that purified HA was present in the void volume fractions, indicating that rHA may form an oligomer. The rHA bound to poly{Neu5Acα2,3LacNAcβ-O[(CH₂)₅NHCO]₂(CH₂)₅NH-/γ-PGA}, which mimics an avian type receptor, but did not bind to γ-polyglutamic acid or human type receptor mimic, poly{Neu5Acα2,6LacNAcβ-O[(CH₂)₅NHCO]₂(CH₂)₅NH-/γ-PGA}, suggesting that it could be utilized as a blocking agent against infection by highly pathogenic influenza viruses.

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“…In both cases, hemolymph containing expressed capsid proteins was used as a crude vaccine solution. In the cabbage looper system, when HAfrom A/PR/8/34 influenza virus (H1N1) expressed in Cabbage looper larvae was immunized into mice using its hemolymph as a crude vaccine solution, anaphylaxis was not observed in the immunized mice [65]. These results suggest that hemolymph that contains subunit vaccines or antigens could be used to vaccinate cattle with an inexpensive formulation.…”

Section: Vlps Production Using Silkworm Larvae and Their Applicationsmentioning

confidence: 99%

“…Moreover, silkworms make the easy and inexpensive scale-up of protein production possible. Several studies have demonstrated that insect larvae [silkworms, T.ni (cabbage looper)] are useful as living biofactories for the inexpensive production of recombinant antigens and vaccines [65]. Insect larvae have also been used for large-scale VLP production.…”

Section: Vlps Production Using Silkworm Larvae and Their Applicationsmentioning

confidence: 99%

Functional Virus-Like Particles Production Using Silkworm and Their Application in Life Science

Kato¹,

Deo²,

Park

et al. 2012

J Biotechnol Biomaterial

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Virus-like particles (VLPs), which mimic the structure of authentic virus, are of significant importance owing to their wide ranging applications. VLPs have the potential to serve as candidate vaccine in addition to subunit vaccines and also serve as a nano-bioparticle scaffold for displaying complex foreign proteins. Here, we review different methods available to produce VLPs with various host expression systems including from microorganisms to mammalian cells and the current potential of silkworms as producers of these VLPs. Recently, silkworms have been used for efficient production of VLPs too in addition to mass production of recombinant proteins, which have contributed to molecular structural analysis, molecule-molecule interactions in biology and biotechnology.

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Insect larvae biofactories as a platform for influenza vaccine production

Cited by 28 publications

References 53 publications

Significant Productivity Improvement of the Baculovirus Expression Vector System by Engineering a Novel Expression Cassette

Significant Productivity Improvement of the Baculovirus Expression Vector System by Engineering a Novel Expression Cassette

Expression and purification of bioactive hemagglutinin protein of highly pathogenic avian influenza A (H5N1) in silkworm larvae

Functional Virus-Like Particles Production Using Silkworm and Their Application in Life Science

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