Effect of osmotic pressure on the production of retroviral vectors: Enhancement in vector stability

Biotechnology and Bioengineering
A S CoroadinhaManuel J T Carrondo

Abstract

The use of Moloney murine leukaemia virus (MoMLV) derived retroviral vectors in gene therapy requires the production of high titer preparations. However, obtaining high titers of infective MoMLV retroviral vectors is difficult due to the vector inherent instability. In this work the effect of the cell culture medium osmotic pressure upon the virus stability was studied. The osmolality of standard medium was raised from 335 up to 500 mOsm/kg using either ionic (sodium chloride) or non-ionic osmotic agents (sorbitol and fructose). It was observed that, independently of the osmotic agent used, the infectious vector inactivation rate was inversely correlated with the osmolality used in the production media; therefore, the use of high medium osmolalities enhanced vector stability. For production purposes a balance must be struck between cell yield, cell productivity and retroviral stability. From the conditions tested herein sorbitol addition, ensuring osmolalities between 410 and 450 mOsm/kg, yields the best production conditions; NaCl hampered the viral infectious production while fructose originates lower cell yields. Lipid extractions were performed for cholesterol and phospholipid analyses showing that more stable viral vectors...Continue Reading

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Citations

Apr 7, 2006·Applied Microbiology and Biotechnology·A S CoroadinhaM J T Carrondo
Jan 19, 2008·Cancer Gene Therapy·C H J LamersJ W Gratama
Apr 18, 2013·Expert Opinion on Biological Therapy·Maria Mercedes SeguraAlain Garnier
Feb 24, 2016·Journal of Chromatography. a·Cláudia S M FernandesA Cecília A Roque
Sep 17, 2008·Journal of Biotechnology·Ana I AmaralPaula M Alves
Aug 20, 2009·Biotechnology Progress·Guan-Yu ChenYu-Chen Hu
Sep 1, 2009·Biotechnology and Bioengineering·Venkata S TayiJames M Piret
Dec 3, 2014·Trends in Biotechnology·Ana F RodriguesAna S Coroadinha
May 17, 2011·Vaccine·Ana Carina SilvaPaula M Alves
Aug 6, 2016·Biotechnology Journal·Cláudia S M FernandesA Cecília A Roque
Jan 22, 2018·Biotechnology Letters·Hugo R SoaresAna S Coroadinha
Sep 1, 2012·Human Gene Therapy Methods·Vanessa BandeiraManuel J T Carrondo
Feb 13, 2021·Viruses·Christopher Perry, Andrea C M E Rayat

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