title: Analytical anion-exchange HPLC of recombinant type-5 adenoviral particles.
abstract: The expanding use of adenoviral vectors for gene therapy has brought about the need for new analytical tools. We have developed an anion-exchange high-performance liquid chromatography method to analyze recombinant adenovirus serotype 5 samples. Before this assay, available analytical methods consisted of either long-term biological assays or required highly purified test articles. These methods were inadequate for optimizing adenovirus production and purification. This assay can quantitate viral particles in either crude lysates or highly pure samples. It can be used to assess particles in both dilute and concentrated samples over a wide dynamic range. Moreover, the population of viral particles eluted in the peak contains most of the infectious virions. This assay is a sensitive technique that overcomes the limitations of previous methods. It provides an essential tool to accomplish process optimization.
authors.affiliation: Canji, Inc., San Diego, CA 92121, USA.
authors.forename: P W
authors.initials: PW
authors.lastname: Shabram
authors.forename: D D
authors.initials: DD
authors.lastname: Giroux
authors.forename: A M
authors.initials: AM
authors.lastname: Goudreau
authors.forename: R J
authors.initials: RJ
authors.lastname: Gregory
authors.forename: M T
authors.initials: MT
authors.lastname: Horn
authors.forename: B G
authors.initials: BG
authors.lastname: Huyghe
authors.forename: X
authors.initials: X
authors.lastname: Liu
authors.forename: M H
authors.initials: MH
authors.lastname: Nunnally
authors.forename: B J
authors.initials: BJ
authors.lastname: Sugarman
authors.forename: S
authors.initials: S
authors.lastname: Sutjipto
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004279:DNA, Viral / Q000302:isolation & purification*; D003673:Defective Viruses / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005069:Evaluation Studies as Topic; D005822:Genetic Vectors; D014461:Ultracentrifugation
chemical_list: D004279:DNA, Viral