Production of GP64-free virus-like particles from baculovirus-infected insect cells

Chaves, Lorena C. S.; Ribeiro, Bergmann Morais; Blissard, Gary W.

doi:10.1099/jgv.0.001002

Cited by 21 publications

(27 citation statements)

References 54 publications

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“…It has been well established that mitigating metabolic limitations during the infection-production phase is more challenging than that for cell growth. This phenomenon referred to as the “cell density effect”, which has been extensively documented by others for insect and human cell production systems infected by viral vectors (Chaves et al 2018; Mlera and Bloom 2019). The specific virus production may drop sharply when infected at a high cell density.…”

Section: Discussionmentioning

confidence: 92%

Development of a simple and high-yielding fed-batch process for the production of porcine circovirus type 2 virus-like particle subunit vaccine

Cao

et al. 2019

AMB Expr

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The cap protein is encoded by the orf2 gene of porcine circovirus type 2 (PCV2) has the main antigen epitope of PCV2 and can form virus-like particles (VLPs), which are expressed in insect cells. PCV2-VLPs can effectively inhibit PCV2 replication as a subunit vaccine. In this study, a robust and reliable fed-batch process was successfully developed for the production of PCV2-VLPs by Sf9 cells. The feeding solution, feeding strategy, and cell density at infection were optimized to maximize the final PCV2-VLPs production yields. The cell density at infection and the volumetric PCV2-VLPs production reached 12 × 106 cells/mL and 110 mg/L, respectively, which yielded 3- and 3.6-fold enhancements compared to the batch culture. The PCV2-VLPs produced in fed-batch culture were not different from the PCV2-VLPs produced in a batch culture in an immunity test. A highly efficient production process was produced for PCV2-VLPs subunit vaccines, which could provide an effective means for the industrial production of PCV2 vaccines.

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Section: Discussionmentioning

confidence: 92%

Development of a simple and high-yielding fed-batch process for the production of porcine circovirus type 2 virus-like particle subunit vaccine

Cao

et al. 2019

AMB Expr

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“…The major limitation of using an insect cell system is the purification step, due to the coproduction of baculovirus particles, a contaminant that must be biophysically or biochemically separated from VLPs (e.g., using cesium chloride ultracentrifugation or ion exchange chromatography), as they can significantly impact production efficiency [8]. This issue can be easily overcome by blocking the formation of baculovirus particles, as has been shown previously in a study of HIV VLP production and purification [36].…”

Section: Discussionmentioning

confidence: 99%

Assembly of tomato blistering mosaic virus-like particles using a baculovirus expression vector system

et al. 2019

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The expression of several structural proteins from a wide variety of viruses in heterologous cell culture systems results in the formation of virus-like particles (VLPs). These VLPs structurally resemble the wild-type virus particles and have been used to study viral assembly process and as antigens for diagnosis and/or vaccine development. Tomato blistering mosaic virus (ToBMV) is a tymovirus that has a 6.3-kb positive-sense ssRNA genome. We have employed the baculovirus expression vector system (BEVS) for the production of tymovirus-like particles (tVLPs) in insect cells. Two recombinant baculoviruses containing the ToBMV wild-type coat protein (CP) gene or a modified short amino-terminal deletion (Δ 2-24 CP) variant were constructed and used to infect insect cells. Both recombinant viruses were able to express ToBMV CP and Δ 2-24 CP from infected insect cells that self-assembled into tVLPs. Therefore, the N-terminal residues (2-24) of the native ToBMV CP were shown not to be essential for self-assembly of tVLPs. We also constructed a third recombinant baculovirus containing a small sequence coding for the major epitope of the chikungunya virus (CHIKV) envelope protein 2 (E2) replacing the native CP N-terminal 2-24 amino acids. This recombinant virus also produced tVLPs. In summary, ToBMV VLPs can be produced in a baculovirus/insect cell heterologous expression system, and the N-terminal residues 2-24 of the CP are not essential for this assembly, allowing its potential use as a protein carrier that facilitates antigen purification and might be used for diagnosis. Handling Editor: Sead Sabanadzovic.

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“…54 In a similar manner, the deletion of GP64, a major baculoviral protein involved in cell entry, 55–57 has been shown to decrease the amount of baculovirus in the supernatant when producing HIV-1 Gag VLP. 58…”

Section: Continuing Efforts To Improve the Baculovirus Expression Vecmentioning

confidence: 99%

The Coming Age of Insect Cells for Manufacturing and Development of Protein Therapeutics

Yee

Zak

Hill

et al. 2018

Ind. Eng. Chem. Res.

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Protein therapeutics is a rapidly growing segment of the pharmaceutical market. Currently, the majority of protein therapeutics are manufactured in mammalian cells for their ability to generate safe and efficacious human-like glycoproteins. The high cost of using mammalian cells for manufacturing has motivated a constant search for alternative host platforms. Insect cells have begun to emerge as a promising candidate, largely due to the development of the baculovirus expression vector system. While there are continuing efforts to improve insect-baculovirus expression for producing protein therapeutics, key limitations including cell lysis and the lack of homogeneous humanized glycosylation still remain. The field has started to see a movement toward virus-less gene expression approaches, notably the use of clustered regularly interspaced short palindromic repeats to address these shortcomings. This review highlights recent technological advances that are realizing the transformative potential of insect cells for the manufacturing and development of protein therapeutics.

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Production of GP64-free virus-like particles from baculovirus-infected insect cells

Cited by 21 publications

References 54 publications

Development of a simple and high-yielding fed-batch process for the production of porcine circovirus type 2 virus-like particle subunit vaccine

Development of a simple and high-yielding fed-batch process for the production of porcine circovirus type 2 virus-like particle subunit vaccine

Assembly of tomato blistering mosaic virus-like particles using a baculovirus expression vector system

The Coming Age of Insect Cells for Manufacturing and Development of Protein Therapeutics

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