Ronald Michalsky - Purification of a recombinant baculovirus of AcMNPV by ion exchange membrane chromatography

Document created by Ronald Michalsky on Aug 22, 2014
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  Publication Details (including relevant citation   information):

  Tanja A.   Grein, Ronald Michalsky, M. Vega Lόpez, Peter Czermak:   Purification of a recombinant baculovirus of AcMNPV by   ion exchange membrane chromatography (submitted,   Journal of Virological Methods).


  Significant progress in the application of viral vectors for gene   delivery into mammalian cells and the use of insect viruses as   sustainable bio pesticides in agriculture require development of   methods for virus purification and concentration. Ion exchange   membrane chromatography is focused in this work as an efficient   and economically attractive tool for virus downstream processing.   Stability of the industrially employed baculovirus Autographa   californica M   nucleopolyhedrovirus was tested at ionic   conductivities of the liquid phase ranging from 0.77-78.00 mS/cm   and at H3O+ activities ranging from pH 3-8.   Viral infectivity rapidly decreased by several orders of   magnitude when ionic conductivities fell below 5 mS/cm (i.e.,   water flux from the surrounding liquid into the viral particle   equivalent to an osmotic pressure change of about 0.49 MPa) or at   H3O+ activities below pH 5.5 (rationalized   with particle aggregation). The virus was concentrated and   purified successfully via adsorption of viral particles to three   different strong anion exchange membranes, in the range of (2.0 ±   1.0) x 109 to (1.1 ± 0.7) x 1010 pfu/ml   membrane, and elution at pH 6.1 and 6.35 mS/cm. Virus recovery   and concentration in accord with the reduction in liquid volume   and concurrent reduction of host cell protein concentrations   below the detection limits were obtained using a polyether   sulfone-based membrane with quaternary ammonium ligand.

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