scholarly journals Production of SARS-CoV-2 virus-like particles in insect cells

2021 ◽  
Author(s):  
Youjun Mi ◽  
Tao Xie ◽  
Bingdong Zhu ◽  
Jiying Tan ◽  
Xuefeng Li ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Human Health ◽  
Insect Cells ◽  
Vaccine Development ◽  
Recombinant Baculovirus ◽  
Expression Plasmid ◽  
Virus Like Particles ◽  
Recombinant Bacmid ◽  
Multiple Cloning Sites ◽  
S Genes

ABSTRACTCoronavirus disease (COVID-19) causes a serious threat to human health. To production of SARS-COV-2 virus-like particles (VLPs) in insect cells for vaccine development and scientific research. The E, M and S genes were cloned into multiple cloning sites of the new triple expression plasmid with one p10 promoter, two pPH promoters and three multiple cloning sites. The plasmid was transformed into DH10 BacTMEscherichia coli competent cells to obtain recombinant bacmid. Then the recombinant bacmid was transfected in ExpiSf9™ insect cells to generate recombinant baculovirus. After ExpiSf9™ infected with the recombinant baculovirus, the E, M, and S protein co-expressed in insect cells. Finally, SARS-CoV-2 VLPs were self-assembled in insect cells after infection. The morphology and the size of SARS-CoV-2 VLPs are similar to the native virions.

scholarly journals Production of SARS-CoV-2 Virus-Like Particles in Insect Cells

Vaccines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 554
Author(s):  
Youjun Mi ◽  
Tao Xie ◽  
Bingdong Zhu ◽  
Jiying Tan ◽  
Xuefeng Li ◽  
...  
Keyword(s):  
Human Health ◽  
Insect Cells ◽  
Vaccine Development ◽  
Recombinant Baculovirus ◽  
Expression Plasmid ◽  
Virus Like Particles ◽  
Competent Cells ◽  
Recombinant Bacmid ◽  
Multiple Cloning Sites ◽  
S Genes

Coronavirus disease (COVID-19) causes a serious threat to human health. Virus-like particles (VLPs) constitute a promising platform in SARS-CoV-2 vaccine development. In this study, the E, M, and S genes were cloned into multiple cloning sites of a new triple expression plasmid with one p10 promoter, two pPH promoters, and three multiple cloning sites. The plasmid was transformed into DH10 BacTMEscherichia coli competent cells to obtain recombinant bacmid. Then the recombinant bacmid was transfected in ExpiSf9TM insect cells to generate recombinant baculovirus. After ExpiSf9TM cells infection with the recombinant baculovirus, the E, M, and S proteins were expressed in insect cells. Finally, SARS-CoV-2 VLPs were self-assembled in insect cells after infection. The morphology and the size of SARS-CoV-2 VLPs are similar to the native virions.

Hepatitis C virus structural proteins and virus-like particles produced in recombinant baculovirus-infected insect cells

2010 ◽  
Vol 44 (1) ◽  
pp. 97-108 ◽  
Author(s):  
S. N. Belzhelarskaya ◽  
N. N. Koroleva ◽  
V. V. Popenko ◽  
V. L. Drutza ◽  
O. V. Orlova ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Insect Cells ◽  
Recombinant Baculovirus ◽  
Structural Proteins ◽  
Virus Like Particles ◽  
Infected Insect

scholarly journals Hepatitis C Virus Structural Proteins Assemble into Viruslike Particles in Insect Cells

1998 ◽  
Vol 72 (5) ◽  
pp. 3827-3836 ◽  
Author(s):  
Thomas F. Baumert ◽  
Susumu Ito ◽  
David T. Wong ◽  
T. Jake Liang
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Insect Cells ◽  
Vaccine Development ◽  
Cultured Cells ◽  
Recombinant Baculovirus ◽  
Structural Proteins ◽  
Particle Assembly ◽  
Viruslike Particles ◽  
Hcv Structural Proteins

ABSTRACT Hepatitis C virus (HCV) is a leading cause of chronic hepatitis in the world. The study of HCV has been hampered by the low level of viral particles in infected individuals, the inability to propagate efficiently the virus in cultured cells, and the lack of a convenient animal model. Due to these obstacles, neither the structure of the virus nor the prerequisites for its assembly have been clearly defined. In this report, we describe a model for the production and purification of HCV-like particles in insect cells using a recombinant baculovirus containing the cDNA of the HCV structural proteins. In insect cells, expressed HCV structural proteins assembled into enveloped viruslike particles (40 to 60 nm in diameter) in large cytoplasmic cisternae, presumably derived from the endoplasmic reticulum. Biophysical characterization of viruslike particles by CsCl and sucrose gradient centrifugation revealed biophysical properties similar to those of putative virions isolated from infected humans. The results suggested that HCV core and envelope proteins without p7 were sufficient for viral particle formation. Analysis of particle-associated nucleic acids demonstrated that HCV RNAs were selectively incorporated into the particles over non-HCV transcripts. The synthesis of HCV-like particles in insect cells may provide an important tool to determine the structural requirements for HCV particle assembly as well as to study viral genome encapsidation and virus-host interactions. The described system may also represent a potential approach toward vaccine development.

scholarly journals Virus-like particles of potato leafroll virus as potential carrier system for nucleic acids.

2005 ◽  
Vol 52 (3) ◽  
pp. 699-702 ◽  
Author(s):  
Elzbieta Sułuja ◽  
Ludmiła Strokowskaja ◽  
Włodzimierz Zagórski-Ostoja ◽  
Andrzej Pałucha
Keyword(s):  
Coat Protein ◽  
Nucleic Acids ◽  
Coat Protein Gene ◽  
Insect Cells ◽  
Protein Gene ◽  
Recombinant Baculovirus ◽  
Potato Leafroll Virus ◽  
Rnase A ◽  
Virus Like Particles ◽  
Leafroll Virus

Potato leafroll virus is a member of the polerovirus genus. The isometric virion is formed by a coat protein encapsidating single-stranded, positive-sense, mono-partite genomic RNA with covalently attached viral protein at the 5' end. The coat protein of the virus exists in two forms: i) a 23 kDa protein, the product of the coat protein gene, and ii) a 78 kDa protein, the product of the coat protein gene and an additional open reading frame expressed by read-through of the coat protein gene stop codon. The aim of this work was the expression of potato leafroll virus coat protein-based proteins that would be able to assemble into virus-like particles in insect cells. These modified particles were tested for their ability to encapsidate nucleic acids. Two types of N-terminally His-tagged coat protein constructs were used for the expression in insect cells: one, encoding a 23 kDa protein with the C-terminal amino-acid sequence corresponding to the wild type coat protein and the second with additional clathrin binding domain at the C-terminus. The expression of these two proteins by a recombinant baculovirus was characterized by Western immunoblotting with antibodies directed against potato leafroll virus. The protection or putative encapsidation of nucleic acids by these two coat protein derivatives was shown by DNase I and RNase A protection assays.

scholarly journals Production of human papillomavirus type 45 virus-like particles in insect cells using a recombinant baculovirus

1996 ◽  
Vol 141 (1) ◽  
pp. 111-116 ◽  
Author(s):  
Antoine Touzé ◽  
Catherine Dupuy ◽  
Martine Chabaud ◽  
Pierre Le Cann ◽  
Pierre Coursaget
Keyword(s):  
Human Papillomavirus ◽  
Insect Cells ◽  
Recombinant Baculovirus ◽  
Virus Like Particles

scholarly journals Recombinant Baculovirus-Produced Grass Carp Reovirus Virus-Like Particles as Vaccine Candidate That Provides Protective Immunity against GCRV Genotype II Infection in Grass Carp

Vaccines ◽  
2021 ◽  
Vol 9 (1) ◽  
pp. 53
Author(s):  
Ting Gao ◽  
Caixia Gao ◽  
Siyu Wu ◽  
Yingying Wang ◽  
Jiyuan Yin ◽  
...  
Keyword(s):  
Grass Carp ◽  
Vaccine Development ◽  
Expression System ◽  
Recombinant Baculovirus ◽  
Ctenopharyngodon Idella ◽  
Economic Losses ◽  
Hemorrhagic Disease ◽  
Grass Carp Reovirus ◽  
Virus Like Particles ◽  
Genotype Ii

Grass carp reovirus (GCRV) leads to severe hemorrhagic disease in grass carp (Ctenopharyngodon idella) and causes economic losses in grass carp aquaculture. Recent epidemiological investigations showed that GCRV genotype II is the dominant subtype in China. Therefore, it is very important to develop a novel vaccine for preventing diseases caused by GCRV genotype II. In this study, we employed a bac-to-bac expression system to generate GCRV-II-based virus-like particles (VLPs). Previous studies have shown that the structural proteins VP3, VP4, and VP38 encoded by the segments S3, S6, and S10 of type II GCRV are immunogenic. Hence, the GCRV-VLPs were produced by co-infection of sf9 cells with recombinant baculoviruses PFBH-VP3, PFBH-VP4, and PFBH-VP38. The expressions of VP3, VP4, and VP38 proteins in GCRV-VLPs were tested by IFA and Western blot analysis. By electron microscopic observations of ultrathin sections, purified VLPs showed that the expressed proteins are similar in shape to GCRV genotype II with a size range from 40 nm to 60 nm. The immunogenicity of GCRV-VLPs was evaluated by the injection immunization of grass carp. The analysis of serum-specific IgM antibody showed that grass carp immunized with GCRV-VLPs produced GCRV-specific antibodies. Furthermore, injection with GCRV-VLPs increased the expressions of immune-related genes (IgM, IFN, TLR3, TLR7) in the spleen and kidney. In addition, grass carp immunized with a GCRV-VLPs-based vaccine showed a relative percent survival rate (RPS) of 83.33% after challenge. The data in this study showed that GCRV-VLPs demonstrated an excellent immunogenicity and represent a promising approach for vaccine development against GCRV genotype II infection.

scholarly journals Immunogenicity and protective potency of Norovirus GII.17 virus-like particle-based vaccine

2018 ◽  
Author(s):  
Wei Chen ◽  
Tao Kang ◽  
Rongliang Yuan ◽  
Yuyang Zhang ◽  
Siqi Xin ◽  
...  
Keyword(s):  
Insect Cells ◽  
Vaccine Development ◽  
Recombinant Baculovirus ◽  
Spherical Particles ◽  
Effective Vaccine ◽  
Viral Gastroenteritis ◽  
Blood Group Antigens ◽  
Electron Microscopic ◽  
Capsid Protein Vp1 ◽  
Norovirus Gii

Noroviruses (NoVs) are a major cause of acute viral gastroenteritis in adults and children worldwide. Lacking of cell culture system and animals models that must be considered the virus like particles (VLPs) used as an effective vaccine development. In the present study, we investigated the expression of the major capsid protein (VP1) of Genogroup II, genotype 17 (GII.17) NoV using recombinant baculovirus system in insect cells and saliva binding blockade assay to detect their protective potency. Our results showed that GII.17 VLPs could be successfully generated in sf9 insect cells and electron microscopic revealed that GII.17 VLPs was visualized as spherical particles of -35nm in diameter. Immunized mouse with purified VLPs produced GII.17 specific sera and could efficiently block GII.17 VLPs binding to saliva histo-blood group antigens (HBGAs). Together, these results suggested that GII.17 VLPs represent a promising vaccine candidate against NoV GII.17 infection and strongly support further preclinical and clinical studies.

Assembly and release of HIV-1 precursor Pr55gag virus-like particles from recombinant baculovirus-infected insect cells

Cell ◽  
1989 ◽  
Vol 59 (1) ◽  
pp. 103-112 ◽  
Author(s):  
Dirk Gheysen ◽  
Eric Jacobs ◽  
Françoise de Foresta ◽  
Clotilde Thiriart ◽  
Myriam Francotte ◽  
...  
Keyword(s):  
Insect Cells ◽  
Recombinant Baculovirus ◽  
Virus Like Particles ◽  
Infected Insect ◽  
Hiv 1
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