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Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System

BACKGROUND: Baculovirus expression system, introduced more than 20 years ago, is considered as a useful tool for large and complex eukaryotic recombinant protein production. A baculovirus expression vector is a recombinant virus which desired foreign protein coding sequences is under control of the...

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Autores principales: Shokrollahi, Narjes, Habibi-Anbouhi, Mahdi, Jahanian-Najafabadi, Ali, Alirahimi, Ehsan, Behdani, Mahdi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Institute of Genetic Engineering and Biotechnology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8217536/
https://www.ncbi.nlm.nih.gov/pubmed/34179196
http://dx.doi.org/10.30498/IJB.2021.2783
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author Shokrollahi, Narjes
Habibi-Anbouhi, Mahdi
Jahanian-Najafabadi, Ali
Alirahimi, Ehsan
Behdani, Mahdi
author_facet Shokrollahi, Narjes
Habibi-Anbouhi, Mahdi
Jahanian-Najafabadi, Ali
Alirahimi, Ehsan
Behdani, Mahdi
author_sort Shokrollahi, Narjes
collection PubMed
description BACKGROUND: Baculovirus expression system, introduced more than 20 years ago, is considered as a useful tool for large and complex eukaryotic recombinant protein production. A baculovirus expression vector is a recombinant virus which desired foreign protein coding sequences is under control of the virus gene promoter. Baculovirus only infects insect cells and do not normally infect vertebrates therefore, they possess no risk of biological risks for human. OBJECTIVES: The aim of this study was to recombinant expression of vascular endothelial growth factor (VEGF) reseptor-2 specific Nanobody in the baculovirus expression system MATERIALS AND METHODS: Gene of specific Nanobody against the VEGF reseptor-2 that called 3VGR19 was cloned and expressed in baculovirus system. RESULTS: 3VGR19 Nanobody gene was amplified by Polymerase Chain Reaction (PCR) using the specific primers, and was cloned in pFastBac HTA plasmid. DH10Bac bacteria was transformed with resulted donor plasmid. The cultured Sf9 insect cell line was transfected with recombinant bacmid, and finally, the expression and purification of 3VGR19 was confirmed in insect cells. CONCLUSIONS: In conclusion, Transient infection of insect cells with baculovirus can be a promising technology for expression of antibody fragments.
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spelling pubmed-82175362021-06-25 Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System Shokrollahi, Narjes Habibi-Anbouhi, Mahdi Jahanian-Najafabadi, Ali Alirahimi, Ehsan Behdani, Mahdi Iran J Biotechnol Research Article BACKGROUND: Baculovirus expression system, introduced more than 20 years ago, is considered as a useful tool for large and complex eukaryotic recombinant protein production. A baculovirus expression vector is a recombinant virus which desired foreign protein coding sequences is under control of the virus gene promoter. Baculovirus only infects insect cells and do not normally infect vertebrates therefore, they possess no risk of biological risks for human. OBJECTIVES: The aim of this study was to recombinant expression of vascular endothelial growth factor (VEGF) reseptor-2 specific Nanobody in the baculovirus expression system MATERIALS AND METHODS: Gene of specific Nanobody against the VEGF reseptor-2 that called 3VGR19 was cloned and expressed in baculovirus system. RESULTS: 3VGR19 Nanobody gene was amplified by Polymerase Chain Reaction (PCR) using the specific primers, and was cloned in pFastBac HTA plasmid. DH10Bac bacteria was transformed with resulted donor plasmid. The cultured Sf9 insect cell line was transfected with recombinant bacmid, and finally, the expression and purification of 3VGR19 was confirmed in insect cells. CONCLUSIONS: In conclusion, Transient infection of insect cells with baculovirus can be a promising technology for expression of antibody fragments. National Institute of Genetic Engineering and Biotechnology 2021-01-01 /pmc/articles/PMC8217536/ /pubmed/34179196 http://dx.doi.org/10.30498/IJB.2021.2783 Text en Copyright: © 2021 The Author(s); Published by Iranian Journal of Biotechnology https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 Unported License, ( http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) ) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Shokrollahi, Narjes
Habibi-Anbouhi, Mahdi
Jahanian-Najafabadi, Ali
Alirahimi, Ehsan
Behdani, Mahdi
Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System
title Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System
title_full Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System
title_fullStr Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System
title_full_unstemmed Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System
title_short Expressing of Recombinant VEGFR2-specific Nanobody in Baculovirus Expression System
title_sort expressing of recombinant vegfr2-specific nanobody in baculovirus expression system
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8217536/
https://www.ncbi.nlm.nih.gov/pubmed/34179196
http://dx.doi.org/10.30498/IJB.2021.2783
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