Scaleable purification of adenovirus vectors.
Adenovirus vectors currently are being evaluated in gene delivery studies ranging from prophylactic vaccination to therapeutic gene therapy. The quantity of purified virus required for these studies necessitate that purification methods must shift from classical density gradient ultracentrifugation to scaleable approaches. A methodology is described herein using batch centrifugation, tangential flow ultrafiltration, and chromatography to purify adenovirus particles at a scale of approximately 10(13) viral particles. This method has been demonstrated to easily scale an additional 40-fold. While purification of human adenovirus type 5 is exemplified, modifications are suggested for the purification of other serotypes.
Konz
John O
JO
Bioprocess R&D, Merck & Co Inc, West Point, PA, USA.
Pitts
Lee R
LR
Sagar
Sangeetha L
SL
D000256:Adenoviridae / Q000235:genetics*; D002460:Cell Line; D002852:Chromatography, Ion Exchange / Q000379:methods*; D005822:Genetic Vectors / Q000302:isolation & purification*; D006801:Humans; D014462:Ultrafiltration