Chaperna-mediated assembly of ferritin-based middle East respiratory syndrome-coronavirus nanoparticles

Young Seok Kim, Ahyun Son, Jihoon Kim, Soon Bin Kwon, Myung Hee Kim, Paul Kim, Jieun Kim, Young Ho Byun, Jemin Sung, Jinhee Lee, Ji Eun Yu, Chan Park, Yeon Sook Kim, Nam Hyuk Cho, Jun Chang, Baik Lin Seong

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The folding of monomeric antigens and their subsequent assembly into higher ordered structures are crucial for robust and effective production of nanoparticle (NP) vaccines in a timely and reproducible manner. Despite significant advances in in silico design and structure-based assembly, most engineered NPs are refractory to soluble expression and fail to assemble as designed, presenting major challenges in the manufacturing process. The failure is due to a lack of understanding of the kinetic pathways and enabling technical platforms to ensure successful folding of the monomer antigens into regular assemblages. Capitalizing on a novel function of RNA as a molecular chaperone (chaperna: chaperone + RNA), we provide a robust protein-folding vehicle that may be implemented to NP assembly in bacterial hosts. The receptor-binding domain (RBD) of Middle East respiratory syndrome-coronavirus (MERS-CoV) was fused with the RNA-interaction domain (RID) and bacterioferritin, and expressed in Escherichia coli in a soluble form. Site-specific proteolytic removal of the RID prompted the assemblage of monomers into NPs, which was confirmed by electron microscopy and dynamic light scattering. The mutations that affected the RNA binding to RBD significantly increased the soluble aggregation into amorphous structures, reducing the overall yield of NPs of a defined size. This underscored the RNA-antigen interactions during NP assembly. The sera after mouse immunization effectively interfered with the binding of MERS-CoV RBD to the cellular receptor hDPP4. The results suggest that RNA-binding controls the overall kinetic network of the antigen folding pathway in favor of enhanced assemblage of NPs into highly regular and immunologically relevant conformations. The concentration of the ion Fe 2+ , salt, and fusion linker also contributed to the assembly in vitro, and the stability of the NPs. The kinetic "pace-keeping" role of chaperna in the super molecular assembly of antigen monomers holds promise for the development and delivery of NPs and virus-like particles as recombinant vaccines and for serological detection of viral infections.

Original languageEnglish
Article number1093
JournalFrontiers in Immunology
Volume9
Issue numberMAY
DOIs
Publication statusPublished - 2018 May 17

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Ferritins
Nanoparticles
RNA
Antigens
Synthetic Vaccines
Molecular Chaperones
Protein Folding
Virus Diseases
Middle East Respiratory Syndrome Coronavirus
Virion
Computer Simulation
Immunization
Electron Microscopy
Vaccines
Salts
Ions
Escherichia coli
Mutation
Serum

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Kim, Young Seok ; Son, Ahyun ; Kim, Jihoon ; Kwon, Soon Bin ; Kim, Myung Hee ; Kim, Paul ; Kim, Jieun ; Byun, Young Ho ; Sung, Jemin ; Lee, Jinhee ; Yu, Ji Eun ; Park, Chan ; Kim, Yeon Sook ; Cho, Nam Hyuk ; Chang, Jun ; Seong, Baik Lin. / Chaperna-mediated assembly of ferritin-based middle East respiratory syndrome-coronavirus nanoparticles. In: Frontiers in Immunology. 2018 ; Vol. 9, No. MAY.
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abstract = "The folding of monomeric antigens and their subsequent assembly into higher ordered structures are crucial for robust and effective production of nanoparticle (NP) vaccines in a timely and reproducible manner. Despite significant advances in in silico design and structure-based assembly, most engineered NPs are refractory to soluble expression and fail to assemble as designed, presenting major challenges in the manufacturing process. The failure is due to a lack of understanding of the kinetic pathways and enabling technical platforms to ensure successful folding of the monomer antigens into regular assemblages. Capitalizing on a novel function of RNA as a molecular chaperone (chaperna: chaperone + RNA), we provide a robust protein-folding vehicle that may be implemented to NP assembly in bacterial hosts. The receptor-binding domain (RBD) of Middle East respiratory syndrome-coronavirus (MERS-CoV) was fused with the RNA-interaction domain (RID) and bacterioferritin, and expressed in Escherichia coli in a soluble form. Site-specific proteolytic removal of the RID prompted the assemblage of monomers into NPs, which was confirmed by electron microscopy and dynamic light scattering. The mutations that affected the RNA binding to RBD significantly increased the soluble aggregation into amorphous structures, reducing the overall yield of NPs of a defined size. This underscored the RNA-antigen interactions during NP assembly. The sera after mouse immunization effectively interfered with the binding of MERS-CoV RBD to the cellular receptor hDPP4. The results suggest that RNA-binding controls the overall kinetic network of the antigen folding pathway in favor of enhanced assemblage of NPs into highly regular and immunologically relevant conformations. The concentration of the ion Fe 2+ , salt, and fusion linker also contributed to the assembly in vitro, and the stability of the NPs. The kinetic {"}pace-keeping{"} role of chaperna in the super molecular assembly of antigen monomers holds promise for the development and delivery of NPs and virus-like particles as recombinant vaccines and for serological detection of viral infections.",
author = "Kim, {Young Seok} and Ahyun Son and Jihoon Kim and Kwon, {Soon Bin} and Kim, {Myung Hee} and Paul Kim and Jieun Kim and Byun, {Young Ho} and Jemin Sung and Jinhee Lee and Yu, {Ji Eun} and Chan Park and Kim, {Yeon Sook} and Cho, {Nam Hyuk} and Jun Chang and Seong, {Baik Lin}",
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Kim, YS, Son, A, Kim, J, Kwon, SB, Kim, MH, Kim, P, Kim, J, Byun, YH, Sung, J, Lee, J, Yu, JE, Park, C, Kim, YS, Cho, NH, Chang, J & Seong, BL 2018, 'Chaperna-mediated assembly of ferritin-based middle East respiratory syndrome-coronavirus nanoparticles', Frontiers in Immunology, vol. 9, no. MAY, 1093. https://doi.org/10.3389/fimmu.2018.01093

Chaperna-mediated assembly of ferritin-based middle East respiratory syndrome-coronavirus nanoparticles. / Kim, Young Seok; Son, Ahyun; Kim, Jihoon; Kwon, Soon Bin; Kim, Myung Hee; Kim, Paul; Kim, Jieun; Byun, Young Ho; Sung, Jemin; Lee, Jinhee; Yu, Ji Eun; Park, Chan; Kim, Yeon Sook; Cho, Nam Hyuk; Chang, Jun; Seong, Baik Lin.

In: Frontiers in Immunology, Vol. 9, No. MAY, 1093, 17.05.2018.

Research output: Contribution to journalArticle

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T1 - Chaperna-mediated assembly of ferritin-based middle East respiratory syndrome-coronavirus nanoparticles

AU - Kim, Young Seok

AU - Son, Ahyun

AU - Kim, Jihoon

AU - Kwon, Soon Bin

AU - Kim, Myung Hee

AU - Kim, Paul

AU - Kim, Jieun

AU - Byun, Young Ho

AU - Sung, Jemin

AU - Lee, Jinhee

AU - Yu, Ji Eun

AU - Park, Chan

AU - Kim, Yeon Sook

AU - Cho, Nam Hyuk

AU - Chang, Jun

AU - Seong, Baik Lin

PY - 2018/5/17

Y1 - 2018/5/17

N2 - The folding of monomeric antigens and their subsequent assembly into higher ordered structures are crucial for robust and effective production of nanoparticle (NP) vaccines in a timely and reproducible manner. Despite significant advances in in silico design and structure-based assembly, most engineered NPs are refractory to soluble expression and fail to assemble as designed, presenting major challenges in the manufacturing process. The failure is due to a lack of understanding of the kinetic pathways and enabling technical platforms to ensure successful folding of the monomer antigens into regular assemblages. Capitalizing on a novel function of RNA as a molecular chaperone (chaperna: chaperone + RNA), we provide a robust protein-folding vehicle that may be implemented to NP assembly in bacterial hosts. The receptor-binding domain (RBD) of Middle East respiratory syndrome-coronavirus (MERS-CoV) was fused with the RNA-interaction domain (RID) and bacterioferritin, and expressed in Escherichia coli in a soluble form. Site-specific proteolytic removal of the RID prompted the assemblage of monomers into NPs, which was confirmed by electron microscopy and dynamic light scattering. The mutations that affected the RNA binding to RBD significantly increased the soluble aggregation into amorphous structures, reducing the overall yield of NPs of a defined size. This underscored the RNA-antigen interactions during NP assembly. The sera after mouse immunization effectively interfered with the binding of MERS-CoV RBD to the cellular receptor hDPP4. The results suggest that RNA-binding controls the overall kinetic network of the antigen folding pathway in favor of enhanced assemblage of NPs into highly regular and immunologically relevant conformations. The concentration of the ion Fe 2+ , salt, and fusion linker also contributed to the assembly in vitro, and the stability of the NPs. The kinetic "pace-keeping" role of chaperna in the super molecular assembly of antigen monomers holds promise for the development and delivery of NPs and virus-like particles as recombinant vaccines and for serological detection of viral infections.

AB - The folding of monomeric antigens and their subsequent assembly into higher ordered structures are crucial for robust and effective production of nanoparticle (NP) vaccines in a timely and reproducible manner. Despite significant advances in in silico design and structure-based assembly, most engineered NPs are refractory to soluble expression and fail to assemble as designed, presenting major challenges in the manufacturing process. The failure is due to a lack of understanding of the kinetic pathways and enabling technical platforms to ensure successful folding of the monomer antigens into regular assemblages. Capitalizing on a novel function of RNA as a molecular chaperone (chaperna: chaperone + RNA), we provide a robust protein-folding vehicle that may be implemented to NP assembly in bacterial hosts. The receptor-binding domain (RBD) of Middle East respiratory syndrome-coronavirus (MERS-CoV) was fused with the RNA-interaction domain (RID) and bacterioferritin, and expressed in Escherichia coli in a soluble form. Site-specific proteolytic removal of the RID prompted the assemblage of monomers into NPs, which was confirmed by electron microscopy and dynamic light scattering. The mutations that affected the RNA binding to RBD significantly increased the soluble aggregation into amorphous structures, reducing the overall yield of NPs of a defined size. This underscored the RNA-antigen interactions during NP assembly. The sera after mouse immunization effectively interfered with the binding of MERS-CoV RBD to the cellular receptor hDPP4. The results suggest that RNA-binding controls the overall kinetic network of the antigen folding pathway in favor of enhanced assemblage of NPs into highly regular and immunologically relevant conformations. The concentration of the ion Fe 2+ , salt, and fusion linker also contributed to the assembly in vitro, and the stability of the NPs. The kinetic "pace-keeping" role of chaperna in the super molecular assembly of antigen monomers holds promise for the development and delivery of NPs and virus-like particles as recombinant vaccines and for serological detection of viral infections.

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