Optimization of chimeric HIV‐1 virus‐like particle production in a baculovirus‐insect cell expression system

Pillay, Sureshnee; Meyers, Ann E.; Williamson, Anna‐Lise; Rybicki, Edward P.

doi:10.1002/btpr.187

Cited by 44 publications

(28 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Due to intrinsic properties of the lipid membrane and surface glycoprotein, the generation of enveloped VLPs in insect cells is more complicated than that of non-enveloped VLPs. However, efficient budding of enveloped VLPs from insect cells has been reported from time to time [30,31,36,38,39,45,46]. For example, using a quadruple baculovirus recombinant, Latham and Galarza (2001) initially showed that co-expression of four structural proteins of influenza virus, the HA, neuraminidase (NA), matrix protein M1 and M2 ion channel protein, was sufficient for the self-assembly and release of VLPs from surface of insect cells.…”

Section: Enveloped Vlpsmentioning

confidence: 99%

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

Liu

et al. 2013

Protein Expression and Purification

106

View full text Add to dashboard Cite

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.

show abstract

Section: Enveloped Vlpsmentioning

confidence: 99%

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

Liu

et al. 2013

Protein Expression and Purification

106

View full text Add to dashboard Cite

show abstract

“…Production of recombinant baculovirus-expressed GagRT and GagTN VLPs was optimized as described in Pillay et al . [16]. VLPs were purified from 2.5 L of Sf 9 cell culture supernatants after 96 h incubation at 27°C.…”

Section: Resultsmentioning

confidence: 99%

HIV-1 sub-type C chimaeric VLPs boost cellular immune responses in mice

Pillay

Shephard

Meyers

et al. 2010

J Immune Based Ther Vaccines

Self Cite

View full text Add to dashboard Cite

Several approaches have been explored to eradicate HIV; however, a multigene vaccine appears to be the best option, given their proven potential to elicit broad, effective responses in animal models. The Pr55Gag protein is an excellent vaccine candidate in its own right, given that it can assemble into large, enveloped, virus-like particles (VLPs) which are highly immunogenic, and can moreover be used as a scaffold for the presentation of other large non-structural HIV antigens. In this study, we evaluated the potential of two novel chimaeric HIV-1 Pr55Gag-based VLP constructs - C-terminal fusions with reverse transcriptase and a Tat::Nef fusion protein, designated GagRT and GagTN respectively - to enhance a cellular response in mice when used as boost components in two types of heterologous prime-boost vaccine strategies. A vaccine regimen consisting of a DNA prime and chimaeric HIV-1 VLP boosts in mice induced strong, broad cellular immune responses at an optimum dose of 100 ng VLPs. The enhanced cellular responses induced by the DNA prime-VLP boost were two- to three-fold greater than two DNA vaccinations. Moreover, a mixture of GagRT and GagTN VLPs also boosted antigen-specific CD8+ and CD4+ T-cell responses, while VLP vaccinations only induced predominantly robust Gag CD4+ T-cell responses. The results demonstrate the promising potential of these chimaeric VLPs as vaccine candidates against HIV-1.

show abstract

“…Virus like particles (VLPs) are multiprotein structures that mimic the characteristics, organization and conformation of native infectious viruses but are themselves noninfectious . These properties have made VLPs an interesting class of molecules for vaccine development More recently, VLPs were introduced as potential BSL‐1‐compatible viral clearance spiking surrogates for biopharmaceutical process development studies .…”

Section: Introductionmentioning

confidence: 99%

Use of a noninfectious surrogate to predict minute virus of mice removal during nanofiltration

Cetlin¹,

Pallansch²,

Fulton³

et al. 2018

Biotechnology Progress

View full text Add to dashboard Cite

Viruses can arise during the manufacture of biopharmaceuticals through contamination or endogenous expression of viral sequences. Regulatory agencies require "viral clearance" validation studies for each biopharmaceutical prior to approval. These studies aim to demonstrate the ability of the manufacturing process at removing or inactivating virus and are conducted by challenging scaled-down manufacturing steps with a "spike" of live virus. Due to the infectious nature of these live viruses, "spiking studies" are typically conducted in specialized biological safety level-2 facilities. The costs and logistics associated with these studies limit viral clearance analysis during process development and characterization. In this study, a noninfectious Minute Virus of Mice-Mock Virus Particle (MVM-MVP) was generated for use as an economical small virus spiking surrogate. An immunoglobin G containing solution was spiked with live MVM or MVM-MVP and processed through Planova nanofiltration units. Flux decay data was collected and particle reduction values were calculated from TCID and Immuno-qPCR analysis. The data indicated comparable filtration performance and particle reduction between infectious MVM and noninfectious surrogate, MVM-MVP. This proof of concept study suggests the feasibility of utilizing MVPs for predictive size-based viral clearance assessments during process development and characterization as an alternative to homologous infectious virus.

show abstract

Optimization of chimeric HIV‐1 virus‐like particle production in a baculovirus‐insect cell expression system

Cited by 44 publications

References 19 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

HIV-1 sub-type C chimaeric VLPs boost cellular immune responses in mice

Use of a noninfectious surrogate to predict minute virus of mice removal during nanofiltration

Contact Info

Product

Resources

About