Human adenoviruses: propagation, purification, quantification, and storage.
Detailed protocols are described for the propagation of adenoviruses (Ads) and adenovirus (Ad) vectors and their purification by CsCl equilibrium density gradient centrifugation. A discussion of monolayer and spinner cell culture techniques suitable, respectively, for small- and large-scale growth of adenoviruses is provided. Protocols for cloning into and growth of Ad replication-deficient vectors using a convenient commercially available system are described. Lastly, time-tested plaque titration protocols for the accurate and convenient measurement of the infectivity of adenoviruses and adenovirus vectors are provided in detail.
Green
Maurice
M
Saint Louis University School of Medicine, St Louis, Missouri, USA.
Loewenstein
Paul M
PM
D000260:Adenoviruses, Human / Q000254:growth & development / Q000302:isolation & purification / Q000502:physiology*; D002499:Centrifugation, Density Gradient; D006801:Humans; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation / Q000379:methods