Conjugation of an antibody Fv fragment to a virus coat protein: cell-specific targeting of recombinant polyoma-virus-like particles

The development of cell-type-specific delivery systems is highly desirable for gene-therapeutic applications. Current virus-based vector systems show broad cell specificity, which results in the need to restrict the natural tropism of these viral systems. Here we demonstrate that tumour-cell-specific virus-like particles can be functionally assembled in vitro from recombinant viral coat protein expressed in Escherichia coli. The insertion of a negatively charged peptide in the HI loop of polyoma VP1 interferes with the binding of VP1 to the natural recognition site on mammalian cells and also serves as an adapter for the coupling of antibody fragments that contain complementary charged fusion peptides. A recombinant antibody fragment of the tumour-specific anti-(Lewis Y) antibody B3 could be coupled to the mutant VP1 by engineered polyionic peptides and an additional disulphide bond. With this system an entirely recombinant cell-specific delivery system assembled in vitro could be generated that transfers genes preferentially to cells presenting the tumour-specific antigen on the cell surface.

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Characteristics of Artificial Virus-like Particles Assembled in vitro from Potato Virus X Coat Protein and Foreign Viral RNAs

Acta Naturae ◽

10.32607/20758251-2011-3-3-40-46 ◽

2011 ◽

Vol 3 (3) ◽

pp. 40-46 ◽

Cited By ~ 8

Author(s):

M V Arkhipenko ◽

E K Petrova ◽

N A Nikitin ◽

A D Protopopova ◽

E V Dubrovin ◽

...

Keyword(s):

Coat Protein ◽

Potato Virus ◽

Potato Virus X ◽

Viral Rnas ◽

Virus Like Particles ◽

Artificial Virus

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Chimeric virus-like particles of universal antigen epitopes of coronavirus and phage Qβ coat protein trigger the production of neutralizing antibodies

Current Topics in Medicinal Chemistry ◽

10.2174/1568026621666210618145411 ◽

2021 ◽

Vol 21 ◽

Author(s):

Yukun Guo ◽

Ruizhen Guo ◽

Yingxian Ma ◽

Wenru Chang ◽

Shengli Ming ◽

...

Keyword(s):

Coat Protein ◽

Neutralizing Antibodies ◽

Vaccine Development ◽

Genetic Recombination ◽

Fluorescent Label ◽

Salting Out ◽

Virus Like Particles ◽

Chimeric Vlps ◽

Universal Epitope

Background: Virus-like particles (VLPs) are non-genetic multimeric nanoparticles synthesized through in vitro or in vivo self-assembly of one or more viral structural proteins. Immunogenicity and safety of VLPs make them ideal candidates for vaccine development and efficient nanocarriers for foreign antigens or adjuvants to activate the immune system. Aims: The present study aimed to design and synthesize a chimeric VLP vaccine of the phage Qbeta (Qβ) coat protein presenting the universal epitope of the coronavirus. Methods: The RNA phage Qβ coat protein was designed and synthesized, denoted as Qbeta. The CoV epitope, a universal epitope of coronavirus, was inserted into the C-terminal of Qbeta using genetic recombination, which was designated as Qbeta-CoV. The N-terminal of Qbeta-CoV was successively inserted into the TEV restriction site using mCherry red fluorescent label and modified affinity-purified histidine label 6xHE, which was denoted as HE-Qbeta-CoV. Isopropyl β-D-1-thiogalactopyranoside (IPTG) assessment revealed the expression of Qbeta, Qbeta-CoV, and HE-Qbeta-CoV in the BL21 (DE3) cells. The fusion protein was purified by salting out using ammonium sulfate and affinity chromatography. The morphology of particles was observed using electron microscopy. The female BALB/C mice were immunized intraperitoneally with the Qbeta-CoV and HE-Qbeta-CoV chimeric VLPs vaccines. Their sera were collected for the detection of antibody level and antibody titer using ELISA. The serum is used for the neutralization test of the three viruses of MHV, PEDV, and PDCoV. Results: The results revealed that the fusion proteins Qbeta, Qbeta-CoV, and HE-Qbeta-CoV could all obtain successful expression. Particles with high purity were obtained after purification; the chimeric particles of Qbeta-CoV and HE-Qbeta-CoV were found to be similar to Qbeta particles in morphology and formed chimeric VLPs. In addition, two chimeric VLP vaccines induced specific antibody responses in mice, and the antibodies showed certain neutralizing activity. Conclusion: The successful construction of the chimeric VLPs of the phage Qβ coat protein presenting the universal epitope of coronavirus provides a vaccine form with potential clinical applications for the treatment of coronavirus disease.

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Hamster polyomavirus-derived virus-like particles are able to transfer in vitro encapsidated plasmid DNA to mammalian cells

Virus Genes ◽

10.1007/s11262-006-0028-1 ◽

2007 ◽

Vol 34 (3) ◽

pp. 303-314 ◽

Cited By ~ 15

Author(s):

Tatyana Voronkova ◽

Andris Kazaks ◽

Velta Ose ◽

Muhsin Özel ◽

Siegfried Scherneck ◽

...

Keyword(s):

Plasmid Dna ◽

Mammalian Cells ◽

Virus Like Particles ◽

Hamster Polyomavirus

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Brome mosaic virus coat protein inhibits viral RNA synthesis in vitro

Virology ◽

10.1016/0042-6822(87)90232-7 ◽

1987 ◽

Vol 158 (1) ◽

pp. 15-19 ◽

Cited By ~ 23

Author(s):

Mamoru Horikoshi ◽

Masaharu Nakayama ◽

Naoto Yamaoka ◽

Iwao Furusawa ◽

Jiko Shishiyama

Keyword(s):

Coat Protein ◽

Mosaic Virus ◽

Rna Synthesis ◽

Brome Mosaic Virus ◽

Viral Rna ◽

Virus Coat Protein ◽

Virus Coat

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Generation and characterization of a recombinant antibody fragment that binds to the coat protein of grapevine leafroll-associated virus 3

Archives of Virology ◽

10.1007/s00705-008-0100-3 ◽

2008 ◽

Vol 153 (6) ◽

pp. 1075-1084 ◽

Cited By ~ 12

Author(s):

Martin Orecchia ◽

Greta Nölke ◽

Pasquale Saldarelli ◽

Mariangela Dell’Orco ◽

Kerstin Uhde-Holzem ◽

...

Keyword(s):

Coat Protein ◽

Recombinant Antibody ◽

Antibody Fragment ◽

Recombinant Antibody Fragment

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Nm23H2 Facilitates Coat Protein Complex II Assembly and Endoplasmic Reticulum Export in Mammalian Cells

Molecular Biology of the Cell ◽

10.1091/mbc.e04-09-0785 ◽

2005 ◽

Vol 16 (2) ◽

pp. 835-848 ◽

Cited By ~ 38

Author(s):

Lori Kapetanovich ◽

Cassandra Baughman ◽

Tina H. Lee

Keyword(s):

Endoplasmic Reticulum ◽

Coat Protein ◽

Protein Complex ◽

Mammalian Cells ◽

Complex Ii ◽

Permeabilized Cells ◽

Er Export ◽

Coat Protein Complex

The cytosolic coat protein complex II (COPII) mediates vesicle formation from the endoplasmic reticulum (ER) and is essential for ER-to-Golgi trafficking. The minimal machinery for COPII assembly is well established. However, additional factors may regulate the process in mammalian cells. Here, a morphological COPII assembly assay using purified COPII proteins and digitonin-permeabilized cells has been applied to demonstrate a role for a novel component of the COPII assembly pathway. The factor was purified and identified by mass spectrometry as Nm23H2, one of eight isoforms of nucleoside diphosphate kinase in mammalian cells. Importantly, recombinant Nm23H2, as well as a catalytically inactive version, promoted COPII assembly in vitro, suggesting a noncatalytic role for Nm23H2. Consistent with a function for Nm23H2 in ER export, Nm23H2 localized to a reticular network that also stained for the ER marker calnexin. Finally, an in vivo role for Nm23H2 in COPII assembly was confirmed by isoform-specific knockdown of Nm23H2 by using short interfering RNA. Knockdown of Nm23H2, but not its most closely related isoform Nm23H1, resulted in diminished COPII assembly at steady state and reduced kinetics of ER export. These results strongly suggest a previously unappreciated role for Nm23H2 in mammalian ER export.

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