Ronald Michalsky - Purification of the baculovirus Autographa californica M nucleopolyhedrovirus by tangential flow ultrafiltration

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    Ronald Michalsky, A. Lorena Passarelli, Peter H. Pfromm, Peter   Czermak: Purification of the baculovirus Autographa   californica M nucleopolyhedrovirus by tangential flow   ultrafiltration, Desalination 245 (2009), p.   694-700.


  Membrane-based virus filtration is used in biotechnological   processes due to advantages such as easy scale-up but is   currently lacking standardized validation protocols. Thus,   ultrafiltration (UF) of a rod-shaped virus was assessed to   contribute to the body of knowledge regarding the   characterization of virus filtration. A recombinant baculovirus   of Autographa californica M nucleopolyhedrovirus (AcMNPV),   vHSGFP, expressing egfp was filtered using polyethersulfone   membranes ranging from 30 to 1000 kDa molecular weight cut-off   (MWCO) and filtration parameters were previously assessed for   their potential to affect virus stability. AcMNPV was   concentrated 20-fold in the retentate using 100-1000 kDa   membranes. However, evidence for a superposition of cake   formation and pore plugging through viruses and other solution   components (likely proteins) causing partially irreversible   fouling of membranes dependent on the MWCO was observed. Results   support UF using a 300 kDa MWCO to concentrate AcMNPV with   minimized membrane fouling.

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