Affinity recovery of Moloney Murine Leukaemia Virus

Lipid enveloped retroviruses such as Moloney Murine Leukaemia Virus (MoMuLV) are commonly used gene therapy vectors. Downstream processing protocols used for their purification are time consuming and a potentially generic, single step capture method for the recovery of retroviral particles is propos...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2005-06, Vol.820 (1), p.111-119
Main Authors: WILLIAMS, Sharon L, NESBETH, Darren, DARLING, David C, FARZANEH, Farzin, SLATER, Nigel K. H
Format: Article
Language:English
Subjects:
Analysis
Analytical, structural and metabolic biochemistry
Animals
Biological and medical sciences
Biotinylation
Chromatography, Affinity - methods
Fundamental and applied biological sciences. Psychology
General pharmacology
HeLa Cells
Humans
Medical sciences
Mice
Moloney murine leukemia virus - chemistry
Moloney murine leukemia virus - isolation & purification
NIH 3T3 Cells
Pharmacology. Drug treatments
Streptavidin
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Summary:Lipid enveloped retroviruses such as Moloney Murine Leukaemia Virus (MoMuLV) are commonly used gene therapy vectors. Downstream processing protocols used for their purification are time consuming and a potentially generic, single step capture method for the recovery of retroviral particles is proposed that exploits streptavidin-biotin affinity chromatography. The ability of four conventional adsorbent solid phases, Fractogel, Sepharose, Magnespheres and STREAMLINE immobilised with streptavidin, to capture and recover biotinylated Moloney Murine Leukaemia Virus was studied. MoMuLV can be biotinylated whilst retaining infectivity and the biotinylated virus can be adsorbed to Streptavidin Magnespheres yielding a 2298-fold increase in titre. For optimal virus biotinylation purification using Fractogel streptavidin can yield a 1896-fold increase in cfu/mg of protein and a 1191-fold decrease in DNA/cfu. Infectious virus can be recovered from Fractogel streptavidin with a maximum recovery of 16.7%.
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2005.03.016