:: Volume 21, Issue 1 (Bimonthly 2017) ::
Feyz 2017, 21(1): 28-34 Back to browse issues page
Evaluation of anticancer peptide VEGF111b secretion in HEK293 human cells
Morteza Sadeghi , Zohreh Hojati
Human Genetic Research Center, Baqiyatallah Medical Sciences University, Tehran, I. R. Iran. , ms.sadeghi@yahoo.com
Abstract:   (3347 Views)

Background: VEGF111b is a new isoform of vascular endothelial growth factor (VEGF) recently considered as a new anticancer drug. The aim of this study was to evaluate the VEGF111b secretion from HEK293 cell wall in order to commercial production of this recombinant factor.

Materials and Methods: After the design of VEGF111b sequence using OLIGO software and NCBI gene bank data, it was cloned into the pBUD.cE4.1 vector. The pBUD.VEGF111b recombinant vector was transfected into HEK293 cells using lipofectamine kit. Forty-eight hours after the transfection the production of VEGF111b was estimated by Western blotting and Human anti VEGF antibody. The VEGF111b secretion   into cell culture and cell lysate extract was measured using ELISA.  

Results: The correct cloning of VEGF111b into pBUD.cE4.1vector was confirmed using enzymatic digestion and gel electrophoresis. The observed production of recombinant peptide in HEK293 was confirmed with 12KDa band in cell lysate extract of Western blotting. The ELISA results at 450 nanometer absorbance for cell culture media and cell lysate extract were 19.20±2.81 pg/ml and 32.87±7.42 pg/ml, respectively. However, no VEGF111b expression was observed in negative controls.

Conclusion: The findings of this study indicate the powerful ability of   transformation and secretion of VEGF111b from HEK293 cell  wall to cell culture media with no breaking and proteolytic digestion. It seems that the commercial production and purification of this therapeutic peptide from HEK293 cell culture would be possible with high efficiency.
Keywords: Vascular Endothelial Growth Factor A, Secretion, HEK293
Full-Text [PDF 338 kb]   (1114 Downloads)    
Type of Study: Research | Subject: General
Received: 2017/02/26 | Revised: 2017/07/19 | Accepted: 2017/02/26 | Published: 2017/02/26
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