Primary recovery and chromatographic purification of adeno-associated virus type 2 produced by baculovirus/insect cell system

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DOIResolve DOI: http://doi.org/10.1016/j.jviromet.2006.09.011
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TypeArticle
Journal titleJournal of Virological Methods
Volume139
Issue1
Pages6170; # of pages: 10
SubjectAnimal; bio; Biotechnology; Cells; Gene Therapy; methods; Spodoptera; therapy; Ultracentrifugation; AAV; Adeno-associated virus; Insect cell; Baculovirus; Purification; Primary recovery
AbstractAdeno-associated virus (AAV) is making its place in gene therapy applications; however, the industry is still facing obstacles in producing a large quantity of highly purified material for clinical studies. Insect cell technology can be used to produce AAV to meet the current demand. During the purification process it was observed that there was a reduced recovery of AAV produced in insect cells, Spodoptera frugiperda (Sf9). It was assumed that the formation of AAV agglomerates and the interaction of AAV with other cellular components were major contributors to this loss. After studying different systems of extraction a sequence of treatment for primary recovery of AAV from cell paste was developed. This sequence was necessary to reduce the AAV losses and to increase the recovery. The purification method avoided the use of ultracentrifugation and adopted chromatographic methods for the purification of AAV. Primary recovery, ion exchange chromatography and hydrophobic interaction chromatography gave an overall yield of 75% from the extracted AAV. The purification process was based on chromatographic methods; therefore, it can be scaled up. Although this method was developed for AAV type 2, it is believed that this method could be modified easily to purify other AAV serotypes.
Publication date
LanguageEnglish
AffiliationNRC Biotechnology Research Institute; National Research Council Canada
Peer reviewedNo
NRC number47782
NPARC number3539375
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Record identifier1f042371-4cee-471e-a35a-535223150ea9
Record created2009-03-01
Record modified2016-05-09
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