Construction and Characterization of Insect Cell-Derived Influenza VLP: Cell Binding, Fusion, and EGFP Incorporation

Pan, Yi-Shin; Wei, Hung-Ju; Chang, Chung‒Chieh; Lin, Chung-Hung; Wei, Sean Ting Shyang; Wu, Suh-Chin; Chang, Ding‐Kwo

doi:10.1155/2010/506363

Cited by 12 publications

(10 citation statements)

References 58 publications

(58 reference statements)

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“…Interestingly, for influenza VLPs, ''removal'' of structurally nonessential glycans on VLPs surface glycoproteins may be a very effective and general approach for VLP-based vaccine design. Truncation of the N-glycan structures on HA can increase sialic acid binding affinities [104], and furthermore these structures are similar to those of insect cell-type N-glycans, which thereby can facilitate the uptake of influenza VLPs by antigen-presenting cells [108]. To date, enveloped influenza VLPs have been developed by biopharmaceutical companies and were demonstrated to induce protective immunity during preclinical and clinical studies [109][110][111][112][113][114][115].…”

Section: Glycosylation Of Recombinant Proteinsmentioning

confidence: 99%

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Liu

et al. 2013

Protein Expression and Purification

104

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The baculovirus expression system (BES) has been one of the versatile platforms for the production of recombinant proteins requiring multiple post-translational modifications, such as folding, oligomerization, phosphorylation, glycosylation, acylation, disulfide bond formation and proteolytic cleavage. Advances in recombinant DNA technology have facilitated application of the BES, and made it possible to express multiple proteins simultaneously in a single infection and to produce multimeric proteins sharing functional similarity with their natural analogs. Therefore, the BES has been used for the production of recombinant proteins and the construction of virus-like particles (VLPs), as well as for the development of subunit vaccines, including VLP-based vaccines. The VLP, which consists of one or more structural proteins but no viral genome, resembles the authentic virion but cannot replicate in cells. The high-quality recombinant protein expression and post-translational modifications obtained with the BES, along with its capacity to produce multiple proteins, imply that it is ideally suited to VLP production. In this article, we critically review the pros and cons of using the BES as a platform to produce both enveloped and non-enveloped VLPs.

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Section: Glycosylation Of Recombinant Proteinsmentioning

confidence: 99%

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Liu

et al. 2013

Protein Expression and Purification

104

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“…A wide range of trackable VLPs has been generated by labeling one or more viral components with a fluorescent protein (27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40). Many VLPs have been labeled after particle production/ purification, usually by chemical coupling to (in)organic Figure 2.…”

Section: Discussionmentioning

confidence: 99%

Generation and characterization of a trackable plant-made influenza H5 virus-like particle (VLP) containing enhanced green fluorescent protein (eGFP)

Young¹,

Arthus-Cartier²,

Yam³

et al. 2015

The FASEB Journal

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Medicago, Inc. has developed an efficient virus-like particle (VLP) vaccine production platform using the Nicotiana benthamiana expression system, and currently has influenza-based products targeting seasonal/pandemic hemagglutinin (HA) proteins in advanced clinical trials. We wished to generate a trackable HA-based VLP that would allow us to study both particle assembly in plants and VLP interactions within the mammalian immune system. To this end, a fusion protein was designed, composed of H5 (from influenza A/Indonesia/05/2005 [H5N1]) with enhanced green fluorescent protein (eGFP). Expression of H5-eGFP in N. benthamiana produced brightly fluorescent ∼160 nm particles resembling H5-VLPs. H5-eGFP-VLPs elicited anti-H5 serologic responses in mice comparable to those elicited by H5-VLPs in almost all assays tested (hemagglutination inhibition/IgG(total)/IgG1/IgG2b/IgG2a:IgG1 ratio), as well as a superior anti-GFP IgG response (mean optical density = 2.52 ± 0.16 sem) to that elicited by soluble GFP (mean optical density = 0.12 ± 0.06 sem). Confocal imaging of N. benthamiana cells expressing H5-eGFP displayed large fluorescent accumulations at the cell periphery, and draining lymph nodes from mice given H5-eGFP-VLPs via footpad injection demonstrated bright fluorescence shortly after administration (10 min), providing proof of concept that the H5-eGFP-protein/VLPs could be used to monitor both VLP assembly and immune trafficking. Given these findings, this novel fluorescent reagent will be a powerful tool to gain further fundamental insight into the biology of influenza VLP vaccines.

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“…All of these insect cell-produced influenza VLPs have been shown to contain uncleaved HA molecules [82]. Binding of influenza VLPs to the cell membrane was shown to result in the VLP-cell fusion (at fusogenic pH) and VLP endocytosis mediated by interaction of HA and the cell sialylated receptor, contributing to VLP antigen presentation [224].…”

Section: Insect Cellsmentioning

confidence: 99%

Virus-like particles as a highly efficient vaccine platform: Diversity of targets and production systems and advances in clinical development

Kushnir

Streatfield

Yusibov

2012

Vaccine

522

403

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Virus-like particles (VLPs) are a class of subunit vaccines that differentiate themselves from soluble recombinant antigens by stronger protective immunogenicity associated with the VLP structure. Like parental viruses, VLPs can be either non-enveloped or enveloped, and they can form following expression of one or several viral structural proteins in a recombinant heterologous system. Depending on the complexity of the VLP, it can be produced in either a prokaryotic or eukaryotic expression system using target-encoding recombinant vectors, or in some cases can be assembled in cell-free conditions. To date, a wide variety of VLP-based candidate vaccines targeting various viral, bacterial, parasitic and fungal pathogens, as well as non-infectious diseases, have been produced in different expression systems. Some VLPs have entered clinical development and a few have been licensed and commercialized. This article reviews VLP-based vaccines produced in different systems, their immunogenicity in animal models and their status in clinical development.

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Construction and Characterization of Insect Cell-Derived Influenza VLP: Cell Binding, Fusion, and EGFP Incorporation

Cited by 12 publications

References 58 publications

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Use of baculovirus expression system for generation of virus-like particles: Successes and challenges

Generation and characterization of a trackable plant-made influenza H5 virus-like particle (VLP) containing enhanced green fluorescent protein (eGFP)

Virus-like particles as a highly efficient vaccine platform: Diversity of targets and production systems and advances in clinical development

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