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Recombinant Expression and Purification of Functional Vascular Endothelial Growth Factor-121 in the Baculovirus Expression System Publisher



Mohseni N1 ; Jahaniannajafabadi A2 ; Kazemilomedasht F1 ; Arezomand R3 ; Habibianbouhi M4 ; Shahbazzadeh D1 ; Behdani M1
Authors
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Authors Affiliations
  1. 1. Biotechnology Research Center, Venom & Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, Iran
  2. 2. Department of Pharmaceutical Biotechnology, School of Pharmacy, Isfahan University of Medical Sciences and Health Services, Isfahan, Iran
  3. 3. Department of Medical Biotechnology and Molecular Science, School of Medicine, North Khorasan University of Medical Science, Bojnurd, Iran
  4. 4. National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran

Source: Asian Pacific Journal of Tropical Medicine Published:2016


Abstract

Objective To express human vascular endothelial growth factor121 (VEGF121) in insect cells. Methods A gene construct containing VEGF was cloned in the pFastBac-HTA vector, followed by transformation in DH10BAC. The recombinant bacmid was then extracted, and transfected into Sf9 insect cells. The transfected cells were harvested, and then VEGF expression was confirmed by western blotting using specific antibodies. The tube formation assay was used for functional assessment of VEGF. Results Our results showed that VEGF could be successfully expressed in the baculovirus system. Purified VEGF was able to stimulate in vitro tube formation of human endothelial cells. Conclusions Results from this study demonstrated that the recombinantly-produced VEGF can be considered as a promising candidate for therapeutic purposes. © 2016 Hainan Medical University
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