Covalent Functionalization of Bioengineered Polyhydroxyalkanoate Spheres Directed by Specific Protein-Protein Interactions

Wong, Jin Xiang; González-Miró, Majela; Sutherland-Smith, Andrew J.; Rehm, Bernd H. A.

doi:10.3389/fbioe.2020.00044

Cited by 20 publications

(33 citation statements)

References 57 publications

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“…BP size, charge and polydispersity were analysed (Fig. 2c ) and were similar to previously produced surface-coated BPs 44 – 49 suggesting structural integrity of BPs and suitability for vaccination. All polydispersity indexes were <35% suggesting suitability for clinical applications 50 .…”

Section: Resultssupporting

confidence: 78%

“…Hence, it is predicted to be suitable for industrial large-scale manufacture and to be significantly more cost-effective than the R21 vaccine, which was produced in yeast. In addition, the BP vaccines are stable at ambient temperature as was previously shown 49 , 60 . Cost-effective scalable manufacture combined with vaccine stability will facilitate dissemination in developing countries where malaria is most relevant.…”

Section: Discussionsupporting

confidence: 66%

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Epitope-coated polymer particles elicit neutralising antibodies against Plasmodium falciparum sporozoites

et al. 2021

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The current Malaria RTS,S vaccine is based on virus-like particles (VLPs) comprising the NANP repetitive epitopes from the cicumsporozoite protein (CSP) of Plasmodium falciparum. This vaccine has limited efficacy, only preventing severe disease in about 30% of vaccinated individuals. A more efficacious vaccine is urgently needed to combat malaria. Here we developed a particulate malaria vaccine based on the same CSP epitopes but using biopolymer particles (BPs) as an antigen carrier system. Specific B- and T-cell epitope-coated BPs were assembled in vivo inside an engineered endotoxin-free mutant of Escherichia coli. A high-yield production process leading to ~27% BP vaccine weight over biomass was established. The epitope-coated BPs were purified and their composition, i.e., the polymer core and epitope identity, was confirmed. Epitope-coated BPs were used alongside soluble peptide epitopes and empty BPs to vaccinate sheep. Epitope-coated BPs showed enhanced immunogenicity by inducing anti-NANP antibody titre of EC50 > 150,000 that were at least 20 times higher than induced by the soluble peptides. We concluded that the additional T-cell epitope was not required as it did not enhance immunogenicity when compared with the B-cell epitope-coated BPs. Antibodies specifically bound to the surface of Plasmodium falciparum sporozoites and efficiently inhibited sporozoite motility and traversal of human hepatocytes. This study demonstrated the utility of biologically self-assembled epitope-coated BPs as an epitope carrier for inclusion in next-generation malaria vaccines.

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Section: Resultssupporting

confidence: 78%

Section: Discussionsupporting

confidence: 66%

Epitope-coated polymer particles elicit neutralising antibodies against Plasmodium falciparum sporozoites

et al. 2021

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“…Anchoring onto a single layer of graphene using SpyTag has been developed for enhancing cryoelectron microscopy structure determination at atomic resolution. 13 SpyTag-linked proteins may be anchored to silica particles assembled through biomimetic silicication 14 or to plastic particles (polyhydroxyalkanoate, PHA) synthesised inside the cell 15 ( Fig. 2A).…”

Section: Application Areas 21 Anchoring To Surfaces or Particlesmentioning

confidence: 99%

Power to the protein: enhancing and combining activities using the Spy toolbox

2020

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“…• Magnetotatic bacteria are difficult to grow-prolonged production time and low production yields ( Wong et al, 2020). Overall, PHA particles seem to provide a versatile platform for in vivo enzyme immobilization, providing competitive advantages over other biological scaffolds ( Table 2).…”

Section: Magnetosomesmentioning

confidence: 99%

Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

Wong

Ogura

Chen

et al. 2020

Front. Bioeng. Biotechnol.

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Enzymes function as biocatalysts and are extensively exploited in industrial applications. Immobilization of enzymes using support materials has been shown to improve enzyme properties, including stability and functionality in extreme conditions and recyclability in biocatalytic processing. This review focuses on the recent advances utilizing the design space of in vivo self-assembled polyhydroxyalkanoate (PHA) particles as biocatalyst immobilization scaffolds. Self-assembly of biologically active enzyme-coated PHA particles is a one-step in vivo production process, which avoids the costly and laborious in vitro chemical cross-linking of purified enzymes to separately produced support materials. The homogeneous orientation of enzymes densely coating PHA particles enhances the accessibility of catalytic sites, improving enzyme function. The PHA particle technology has been developed into a remarkable scaffolding platform for the design of cost-effective designer biocatalysts amenable toward robust industrial bioprocessing. In this review, the PHA particle technology will be compared to other biological supramolecular assembly-based technologies suitable for in vivo enzyme immobilization. Recent progress in the fabrication of biological particulate scaffolds using enzymes of industrial interest will be summarized. Additionally, we outline innovative approaches to overcome limitations of in vivo assembled PHA particles to enable finetuned immobilization of multiple enzymes to enhance performance in multi-step cascade reactions, such as those used in continuous flow bioprocessing.

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Covalent Functionalization of Bioengineered Polyhydroxyalkanoate Spheres Directed by Specific Protein-Protein Interactions

Cited by 20 publications

References 57 publications

Epitope-coated polymer particles elicit neutralising antibodies against Plasmodium falciparum sporozoites

Epitope-coated polymer particles elicit neutralising antibodies against Plasmodium falciparum sporozoites

Power to the protein: enhancing and combining activities using the Spy toolbox

Bioengineered Polyhydroxyalkanoates as Immobilized Enzyme Scaffolds for Industrial Applications

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