Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli

Macrobrachium rosenbergii nodavirus (MrNV) is a virus native to giant freshwater prawn. Recombinant MrNV capsid protein has been produced in Escherichia coli, which self-assembled into virus-like particles (VLPs). However, this recombinant protein is unstable, degrading and forming heterogenous VLPs...

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Main Authors: Kueh, Chare Li, Yong, Chean Yeah, Dezfooli, Seyedehsara Masoomi, Bhassu, Subha, Tan, Soon Guan, Tan, Wen Siang
Format: Article
Language:English
Published: American Institute of Chemical Engineers 2017
Online Access:http://psasir.upm.edu.my/id/eprint/59658/
http://psasir.upm.edu.my/id/eprint/59658/1/Virus-like%20particle%20of%20Macrobrachium%20rosenbergii%20nodavirus%20produced%20in%20Spodoptera%20frugiperda%20%28Sf9%29%20cells%20is%20distinctive%20from%20that%20produced%20in%20Escherichia%20coli.pdf
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author Kueh, Chare Li
Yong, Chean Yeah
Dezfooli, Seyedehsara Masoomi
Bhassu, Subha
Tan, Soon Guan
Tan, Wen Siang
author_facet Kueh, Chare Li
Yong, Chean Yeah
Dezfooli, Seyedehsara Masoomi
Bhassu, Subha
Tan, Soon Guan
Tan, Wen Siang
author_sort Kueh, Chare Li
building UPM Institutional Repository
collection Online Access
description Macrobrachium rosenbergii nodavirus (MrNV) is a virus native to giant freshwater prawn. Recombinant MrNV capsid protein has been produced in Escherichia coli, which self-assembled into virus-like particles (VLPs). However, this recombinant protein is unstable, degrading and forming heterogenous VLPs. In this study, MrNV capsid protein was produced in insect Spodoptera frugiperda (Sf9) cells through a baculovirus system. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed that the recombinant protein produced by the insect cells self-assembled into highly stable, homogenous VLPs each of approximately 40 nm in diameter. Enzyme-linked immunosorbent assay (ELISA) showed that the VLPs produced in Sf9 cells were highly antigenic and comparable to those produced in E. coli. In addition, the Sf9 produced VLPs were highly stable across a wide pH range (2–12). Interestingly, the Sf9 produced VLPs contained DNA of approximately 48 kilo base pairs and RNA molecules. This study is the first report on the production and characterization of MrNV VLPs produced in a eukaryotic system. The MrNV VLPs produced in Sf9 cells were about 10 nm bigger and had a uniform morphology compared with the VLPs produced in E. coli. The insect cell production system provides a good source of MrNV VLPs for structural and immunological studies as well as for host–pathogen interaction studies.
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spelling upm-596582018-03-15T03:17:56Z http://psasir.upm.edu.my/id/eprint/59658/ Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli Kueh, Chare Li Yong, Chean Yeah Dezfooli, Seyedehsara Masoomi Bhassu, Subha Tan, Soon Guan Tan, Wen Siang Macrobrachium rosenbergii nodavirus (MrNV) is a virus native to giant freshwater prawn. Recombinant MrNV capsid protein has been produced in Escherichia coli, which self-assembled into virus-like particles (VLPs). However, this recombinant protein is unstable, degrading and forming heterogenous VLPs. In this study, MrNV capsid protein was produced in insect Spodoptera frugiperda (Sf9) cells through a baculovirus system. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) revealed that the recombinant protein produced by the insect cells self-assembled into highly stable, homogenous VLPs each of approximately 40 nm in diameter. Enzyme-linked immunosorbent assay (ELISA) showed that the VLPs produced in Sf9 cells were highly antigenic and comparable to those produced in E. coli. In addition, the Sf9 produced VLPs were highly stable across a wide pH range (2–12). Interestingly, the Sf9 produced VLPs contained DNA of approximately 48 kilo base pairs and RNA molecules. This study is the first report on the production and characterization of MrNV VLPs produced in a eukaryotic system. The MrNV VLPs produced in Sf9 cells were about 10 nm bigger and had a uniform morphology compared with the VLPs produced in E. coli. The insect cell production system provides a good source of MrNV VLPs for structural and immunological studies as well as for host–pathogen interaction studies. American Institute of Chemical Engineers 2017 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/59658/1/Virus-like%20particle%20of%20Macrobrachium%20rosenbergii%20nodavirus%20produced%20in%20Spodoptera%20frugiperda%20%28Sf9%29%20cells%20is%20distinctive%20from%20that%20produced%20in%20Escherichia%20coli.pdf Kueh, Chare Li and Yong, Chean Yeah and Dezfooli, Seyedehsara Masoomi and Bhassu, Subha and Tan, Soon Guan and Tan, Wen Siang (2017) Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli. Biotechnology Progress, 33 (2). pp. 549-557. ISSN 8756-7938; ESSN: 1520-6033 http://onlinelibrary.wiley.com/wol1/doi/10.1002/btpr.2409/abstract 10.1002/btpr.2409
spellingShingle Kueh, Chare Li
Yong, Chean Yeah
Dezfooli, Seyedehsara Masoomi
Bhassu, Subha
Tan, Soon Guan
Tan, Wen Siang
Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli
title Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli
title_full Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli
title_fullStr Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli
title_full_unstemmed Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli
title_short Virus-like particle of Macrobrachium rosenbergii nodavirus produced in Spodoptera frugiperda (Sf9) cells is distinctive from that produced in Escherichia coli
title_sort virus-like particle of macrobrachium rosenbergii nodavirus produced in spodoptera frugiperda (sf9) cells is distinctive from that produced in escherichia coli
url http://psasir.upm.edu.my/id/eprint/59658/
http://psasir.upm.edu.my/id/eprint/59658/
http://psasir.upm.edu.my/id/eprint/59658/
http://psasir.upm.edu.my/id/eprint/59658/1/Virus-like%20particle%20of%20Macrobrachium%20rosenbergii%20nodavirus%20produced%20in%20Spodoptera%20frugiperda%20%28Sf9%29%20cells%20is%20distinctive%20from%20that%20produced%20in%20Escherichia%20coli.pdf