Structure-based antigen design: a strategy for next generation vaccines

Dormitzer, Philip R.; Ulmer, Jeffrey B.; Rappuoli, Rino

doi:10.1016/j.tibtech.2008.08.002

Cited by 153 publications

(123 citation statements)

References 69 publications

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“…The study by Connors et al identified a similarlylocated 37-kDa band as a degraded VP1 fragment, designated VP1*, which resulted from secondary thrombin proteolysis at Arg58 of VP1 . A similar digestion profile was observed for VP1-H3-H190-H190 (lane [4][5][6]. Three cleavage products were observed for the modular construct: (i) modular VP1 (bands at about 47 kDa; blue arrow), (ii) a GST tag (bands at about 25 kDa; orange arrow), and (iii) a cleavage product, which was represented as bands at approximately 39 kDa (green arrow).…”

Section: Sds-pagesupporting

confidence: 66%

“…Furthermore, the SDS-PAGE analysis showed that a small proportion of the 70-kDa contaminant (orange arrow) was detected in the earlier fractions of the eluted peak, which were fractions F1-F3 (lanes [4][5][6]. The proportion of the 70-kDa contaminant was considerably smaller than the proportion of the contaminant in the SEC-purified (lane 1)…”

Section: In Bed Modementioning

confidence: 96%

“…Fusion of different protein domains to realize a synergic combination of modules [3] has been used extensively to create hybrid enzymes [4] and humanized antibodies [5]. This approach has yet to impact vaccines significantly, but rational vaccine design based on presentation of protective antigen modules [6,7] is emerging.…”

Section: Introductionmentioning

confidence: 99%

“…. Figure [6][7]. SDS-PAGE analysis of GST-tag removal from modular VP1 and wt-VP1 (wt-VP1 residues 1-384) using thrombin.…”

mentioning

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: Sds-pagesupporting

confidence: 66%

Section: In Bed Modementioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

“…. Figure [6][7]. SDS-PAGE analysis of GST-tag removal from modular VP1 and wt-VP1 (wt-VP1 residues 1-384) using thrombin.…”

mentioning

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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“…This approach to vaccination has been termed "rational" or "structural" vaccination since the 89 . In the case of anthrax, PA requires optimization for improved storage and a well-defined vaccine product that elicits longer-lasting immunity with fewer doses.…”

Section: Rationally Designed Vaccinesmentioning

confidence: 99%

Design and Testing of Novel Anthrax Vaccines Utilizing a Tobacco Mosaic Virus Expression System

McComb¹

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Structure‐based design of ferritin nanoparticle immunogens displaying antigenic loops of Neisseria gonorrhoeae

et al. 2017

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Effective vaccines are urgently needed to combat gonorrhea, a common sexually transmitted bacterial infection, for which treatment options are diminishing due to rapid emergence of antibiotic resistance. We have used a rational approach to the development of gonorrhea vaccines, and genetically engineered nanoparticles to present antigenic peptides of Neisseria gonorrhoeae , the causative agent of gonorrhea. We hypothesized that the ferritin nanocage could be used as a platform to display an ordered array of N. gonorrhoeae antigenic peptides on its surface. MtrE, the outer membrane channel of the highly conserved gonococcal MtrCDE active efflux pump, is an attractive vaccine target due to its importance in protecting N. gonorrhoeae from host innate effectors and antibiotic resistance. Using computational approaches, we designed constructs that expressed chimeric proteins of the Helicobacter pylori ferritin and antigenic peptides that correspond to the two surface‐exposed loops of N. gonorrhoeae MtrE. The peptides were inserted at the N terminus or in a surface‐exposed ferritin loop between helices αA and αB. Crystal structures of the chimeric proteins revealed that the proteins assembled correctly into a 24‐mer nanocage structure. Although the inserted N. gonorrhoeae peptides were disordered, it was clear that they were displayed on the nanocage surface, but with multiple conformations. Our results confirmed that the ferritin nanoparticle is a robust platform to present antigenic peptides and therefore an ideal system for rational design of immunogens.

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Structure-based antigen design: a strategy for next generation vaccines

Cited by 153 publications

References 69 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

Design and Testing of Novel Anthrax Vaccines Utilizing a Tobacco Mosaic Virus Expression System

Structure‐based design of ferritin nanoparticle immunogens displaying antigenic loops of Neisseria gonorrhoeae

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