A microbial platform for rapid and low-cost virus-like particle and capsomere vaccines

Middelberg, Anton P. J.; Rivera-Hernandez, Tania; Wibowo, Nani; Lua, Linda H.L.; Fan, Yu; Magor, Graham; Chang, Cindy J.; Chuan, Yap P.; Good, Michael F.; Batzloff, Michael R.

doi:10.1016/j.vaccine.2011.05.075

Cited by 86 publications

(170 citation statements)

References 63 publications

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“…coli codon-optimized DNA oligomers corresponding to the antigen modules containing heterologous elements (Fig. 1B) were cloned into pGEX-VP1 vectors as described previously [9]. Modular proteins VP1-GCN4-H190-GCN4 and VP1-H190-H190 were recombinantly produced and prepared as described [45,46].…”

Section: Cloning Purification and Assembly Of Modular Vlpsmentioning

confidence: 99%

“…Modular proteins VP1-GCN4-H190-GCN4 and VP1-H190-H190 were recombinantly produced and prepared as described [45,46]. After endotoxin removal to below 5 EU/ml, purified modular capsomeres were assembled in vitro into modular VLPs and dialyzed against PBS [9]. The endotoxin levels of the capsomeres were below the suggested limit for a recombinant protein based vaccine, which is 20 EU/ml [47].…”

Section: Cloning Purification and Assembly Of Modular Vlpsmentioning

confidence: 99%

“…VLPs are protein assemblies that mimic viral structure [8][9][10][11][12][13][14]; modular VLPs combine a VLP with an antigen module from an unrelated pathogen [9,[15][16][17]. Two types of modules are commonly used: (i) short peptides expected to have minimal structure [18][19][20][21]; and (ii) larger domains with expected tertiary structures [22][23][24][25].…”

Section: Introductionmentioning

confidence: 99%

“…High-efficiency microbial expression and purification processes for modular MuPyV VLPs have been reported [9]. We explore whether the helicity of different H190 antigen module designs affect the quality, i.e.…”

Section: Introductionmentioning

confidence: 99%

“…GCN4 is well characterized and has been used as an ␣-helical scaffold [35,36] or as a helix promoter [37,38]. We previously reported the inclusion of GCN4 in an antigenic module designed to display Group A Streptococcal antigen on a VLP [9]. However, these studies have not established whether the GCN4 elements have any positive effect on the immune response quality.…”

Section: Introductionmentioning

confidence: 99%

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Engineering of virus-like particles for alternative vaccine candidate targeting a hypervariable peptide antigen element

Anggraeni¹

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A promising alternative to replace the current egg-or cell culture-based technology for vaccine production from live viruses is virus-like particle (VLP) technology based on a microbial platform. VLPs are macromolecular assemblies of viral capsid proteins, which have been shown to tolerate insertion of antigen modules via genetic recombinant technology, yielding modular VLPs. Many studies on modular VLPs presume that when a peptide antigen element is taken out from the intact proteins and then modularised on VLPs, it is unable to fold into its native structure. However, until now, presentation of a peptide antigen element on a VLP and the impact of the display strategy to present the antigen element on the quality of the resulting antibodies (i.e. the ability of the antibodies to recognise the intact protein) are not fully understood. This thesis aims to understand the underlying fundamentals regarding modularisation of peptide antigen elements on VLPs for induction of high-quality antibodies. A hypervariable receptor-binding domain, Helix 190 (H190), from the hemagglutinin protein of influenza A virus was used as a model for modularisation on VLPs from murine polyomavirus (MuPyV) VP1 protein. Four major findings are presented. Firstly, two display strategies, i.e. arraying of H190 in tandem repeats and the use of helix promoter elements, were shown to display H190 in its immunogenic form equally. However, modularisation using tandem repeat display induced antibodies of a higher quality than modularisation using helix promoter elements.Secondly, the quality of antibodies induced by the tandem repeat display bearing two copies of H190 was optimum, thus no significant improvement was observed following the use of adjuvant or increasing the copy number of H190. Additionally, the increase in the copy number of H190 was shown to reduce the assembly capability and solubility of modular VP1 in an environment that was optimised for wild-type VP1. Thirdly, this thesis shows the novel finding in the use of flanking ionic elements to stabilise VLP precursors, termed as capsomeres, bearing two copies of H190 containing a hydrophobic stretch, which caused aggregation. Fourthly, the first steps towards obtaining the atomic crystal structure of presented H190 on a modular protein were performed, i.e. a mild and satisfactory laboratory process was developed to achieve high-purity modular VP1 capsomeres, unattainable using previously established expression and purification process of wild-type MuPyV VP1. This thesis shows a step forward towards understanding the presentation of a peptide antigen element on a VLP that enables induction of highquality antibodies, and towards VLP engineering to manipulate the aggregation and solubility of modular VP1. VLP technology based on a microbial platform presented here is iii a potentially safe and effective alternative vaccine candidate that targets a hypervariable peptide antigen element. The speed of the microbial platform allows a rapid response to the hypervariability of the pe...

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Section: Cloning Purification and Assembly Of Modular Vlpsmentioning

confidence: 99%

Section: Cloning Purification and Assembly Of Modular Vlpsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Engineering of virus-like particles for alternative vaccine candidate targeting a hypervariable peptide antigen element

Anggraeni¹

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Nanoscience versus Viruses: The SARS‐CoV‐2 Case

Gościańska

Freund

Wuttke

2021

Adv Funct Materials

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The outbreak of the coronavirus (SARS‐CoV‐2) pandemic, its rapid spread, and its fatal consequences clearly showed mankind the overwhelming power of nature, and the importance of interdisciplinary research to counter it. Herein, it will be pointed out what challenges arise in fighting nanosized viruses, which feature an outstanding ability to alter their structure and properties to adapt to infection processes, and why nanoscience is extremely powerful to address them. To learn from the past and to be best prepared for the future, this review highlights how the incredible potential of nanoscience can be tapped by mimicking natures’ viruses for the development of maximally efficient pharmaceuticals, e.g., COVID‐19 vaccines, by designing preventive and protective equipment, e.g., face masks, as well as developing nanosensors for early detection of infections.

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Different applications of virus‐like particles in biology and medicine: Vaccination and delivery systems

2015

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Virus-like particles (VLPs) mimic the whole construct of virus particles devoid of viral genome as used in subunit vaccine design. VLPs can elicit efficient protective immunity as direct immunogens compared to soluble antigens co-administered with adjuvants in several booster injections. Up to now, several prokaryotic and eukaryotic systems such as insect, yeast, plant, and E. coli were used to express recombinant proteins, especially for VLP production. Recent studies are also generating VLPs in plants using different transient expression vectors for edible vaccines. VLPs and viral particles have been applied for different functions such as gene therapy, vaccination, nanotechnology, and diagnostics. Herein, we describe VLP production in different systems as well as its applications in biology and medicine.

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A microbial platform for rapid and low-cost virus-like particle and capsomere vaccines

Cited by 86 publications

References 63 publications

Engineering of virus-like particles for alternative vaccine candidate targeting a hypervariable peptide antigen element

Engineering of virus-like particles for alternative vaccine candidate targeting a hypervariable peptide antigen element

Nanoscience versus Viruses: The SARS‐CoV‐2 Case

Different applications of virus‐like particles in biology and medicine: Vaccination and delivery systems

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