Production of HIV-1-based virus-like particles for vaccination: achievements and limits

Cervera, Laura; Gòdia, Francesc; Tarrés-Freixas, Ferran; Aguilar‐Gurrieri, Carmen; Carrillo, Jorge; Blanco, Julià; Gutiérrez-Granados, Sònia

doi:10.1007/s00253-019-10038-3

Cited by 38 publications

(68 citation statements)

References 151 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Their success as immunogens lies on their ability to mimic native viruses without containing a viral genome. Their highly organized and repetitive antigen structure has shown effective cellular and humoral immune responses [2]. Furthermore, advances in the field of bioengineering have widened their possible applications; VLP technology accepts several modifications including encapsulation, chemical conjugation or genetic engineering.…”

Section: Introductionmentioning

confidence: 99%

“…The study of PSD, particle ultrastructural analysis, VLP quantification and differentiation from other nanoparticle subpopulations has been performed. The VLPs analyzed in this work are HIV-1 Gag VLPs, which are a promising platform for the development of a vaccine candidate against HIV, but also as a scaffold for chimeric or multivalent vaccine development [2,35]. Upon expression in the host cell, the Gag polyprotein travels to the cell membrane and after an oligomerization process, HIV-1 Gag VLPs are released from the cell through a budding process [36,37].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Quality Assessment of Virus-Like Particles at Single Particle Level: A Comparative Study

Puente‐Massaguer

Cervera

Gòdia

2020

Viruses

Self Cite

View full text Add to dashboard Cite

Virus-like particles (VLPs) have emerged as a powerful scaffold for antigen presentation and delivery strategies. Compared to single protein-based therapeutics, quality assessment requires a higher degree of refinement due to the structure of VLPs and their similar properties to extracellular vesicles (EVs). Advances in the field of nanotechnology with single particle and high-resolution analysis techniques provide appealing approaches to VLP characterization. In this study, six different biophysical methods have been assessed for the characterization of HIV-1-based VLPs produced in mammalian and insect cell platforms. Sample preparation and equipment set-up were optimized for the six strategies evaluated. Electron Microscopy (EM) disclosed the presence of several types of EVs within VLP preparations and cryogenic transmission electron microscopy (cryo-TEM) resulted in the best technique to resolve the VLP ultrastructure. The use of super-resolution fluorescence microscopy (SRFM), nanoparticle tracking analysis (NTA) and flow virometry enabled the high throughput quantification of VLPs. Interestingly, differences in the determination of nanoparticle concentration were observed between techniques. Moreover, NTA and flow virometry allowed the quantification of both EVs and VLPs within the same experiment while analyzing particle size distribution (PSD), simultaneously. These results provide new insights into the use of different analytical tools to monitor the production of nanoparticle-based biologicals and their associated contaminants.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Quality Assessment of Virus-Like Particles at Single Particle Level: A Comparative Study

Puente‐Massaguer

Cervera

Gòdia

2020

Viruses

Self Cite

View full text Add to dashboard Cite

show abstract

“…Their success as immunogens and nanocarriers has been demonstrated in numerous preclinical and clinical trials over the last years (Charlton Hume et al, 2019). Among the different VLP candidates, enveloped human immunodeficiency virus‐1 (HIV‐1) Gag VLPs are in the spotlight as a scaffold for antigen presentation against different diseases (Cervera et al, 2019; Charlton Hume et al, 2019) and as a reference material for the production of extracellular vesicles (EVs; Geeurickx et al, 2019). The HIV‐1 structural polyprotein (Gag) has the capacity to form noninfective viral particles on their own (Göttlinger, 2001).…”

Section: Introductionmentioning

confidence: 99%

“…These production platforms are widely used in industry, and have been brought to the production scale, achieving competitive production yields for preclinical studies (Cruz et al, 1998; Fuenmayor, Cervera, Gòdia, & Kamen, 2019; Venereo‐Sanchez et al, 2017). Although transiently produced VLPs in mammalian cells show lower productivities, the BEVS present a major bottleneck in product purity (Cervera et al, 2019). In fact, co‐purification between Gag VLPs and baculoviruses is of general concern, owing to their similar physicochemical and biochemical properties (Wang et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Quantification of the HIV‐1 virus‐like particle production process by super‐resolution imaging: From VLP budding to nanoparticle analysis

Puente‐Massaguer

Cervera

Gòdia

2020

Biotech & Bioengineering

Self Cite

View full text Add to dashboard Cite

Virus‐like particles (VLPs) offer great promise in the field of nanomedicine. Enveloped VLPs are a class of these nanoparticles and their production process occurs by a budding process, which is known to be the most critical step at intracellular level. In this study, we developed a novel imaging method based on super‐resolution fluorescence microscopy (SRFM) to assess the generation of VLPs in living cells. This methodology was applied to study the production of Gag VLPs in three animal cell platforms of reference: HEK 293‐transient gene expression (TGE), High Five‐baculovirus expression vector system (BEVS) and Sf9‐BEVS. Quantification of the number of VLP assembly sites per cell ranged from 500 to 3,000 in the different systems evaluated. Although the BEVS was superior in terms of Gag polyprotein expression, the HEK 293‐TGE platform was more efficient regarding the assembly of Gag as VLPs. This was translated into higher levels of non‐assembled Gag monomer in BEVS harvested supernatants. Furthermore, the presence of contaminating nanoparticles was evidenced in all three systems, specifically in High Five cells. The SRFM‐based method here developed was also successfully applied to measure the concentration of VLPs in crude supernatants. The lipid membrane of VLPs and the presence of nucleic acids alongside these nanoparticles could also be detected using common staining procedures. Overall, a complete picture of the VLP production process was achieved in these three production platforms. The robustness and sensitivity of this new approach broaden the applicability of SRFM toward the development of new detection, diagnosis and quantification methods based on confocal microscopy in living systems.

show abstract

“…An approach to the development of artificial proteins constructed using conservative T-and B-cell epitopes and their mimics is also promising for the development of vaccines against different viruses, including HIV [7,28]. The synthesis of vaccines against HIV-1 using virus-like nanoparticles is another area of research in HIV vaccines [29][30][31][32].…”

Section: Hiv Vaccine Candidates and Antibodies Expressed In Plantsmentioning

confidence: 99%

Plant-made HIV vaccines and potential candidates

Tremouillaux-Guiller,

Moustafa,

Hefferon

et al. 2020

Preprint

View full text Add to dashboard Cite

Millions of people around the world suffer from heavy social and health burdens related to HIV/AIDS and its associated opportunistic infections. To reduce these burdens, preventive and therapeutic vaccines are required. Effective HIV vaccines have been under investigation for several decades using different animal models. Potential plant-made HIV vaccine candidates have also gained attention in the past few years. In addition to this, broadly neutralizing antibodies produced in plants which can target conserved viral epitopes and neutralize mutating HIV strains have been identified. Numerous epitopes of glycoproteins and capsid proteins of HIV-1 are a part of HIV therapy. Here, we discuss some recent findings aiming to produce anti-HIV-1 recombinant proteins in engineered plants for AIDS prophylactics and therapeutic treatments.

show abstract

Production of HIV-1-based virus-like particles for vaccination: achievements and limits

Cited by 38 publications

References 151 publications

Quality Assessment of Virus-Like Particles at Single Particle Level: A Comparative Study

Quality Assessment of Virus-Like Particles at Single Particle Level: A Comparative Study

Quantification of the HIV‐1 virus‐like particle production process by super‐resolution imaging: From VLP budding to nanoparticle analysis

Plant-made HIV vaccines and potential candidates

Contact Info

Product

Resources

About