Rescue, amplification, purification, and PEGylation of replication defective first-generation adenoviral vectors.
Adenoviral gene therapy vectors have been widely studied and used. Their extremely high transduction efficiency and gene delivery in vivo make them attractive for cancer gene therapy approaches. While they are robust, they are also very immunogenic. One approach to mitigate the immunogenicity of adenoviruses and to evade neutralizing antibodies is to coat the virus with the hydrophilic polymer polyethylene glycol (PEG) (1). This chapter details the steps involved when going from recombinant adenoviral vector plasmid all the way to validated PEGylated adenovirus product.
Barry
Michael A
MA
Mayo Clinic, Rochester, MN, USA.
Weaver
Eric A
EA
Hofherr
Sean E
SE
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology*; D000588:Amines / Q000032:analysis; D001681:Biological Assay; D002021:Buffers; D002586:Cesium; D002712:Chlorides; D005822:Genetic Vectors / Q000235:genetics* / Q000302:isolation & purification*; D016679:Genome, Viral / Q000235:genetics; D008027:Light; D008156:Luciferases / Q000378:metabolism; D010316:Particle Size; D011092:Polyethylene Glycols / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012542:Scattering, Radiation; D014161:Transduction, Genetic; D014779:Virus Replication / Q000502:physiology*
D000588:Amines; D002021:Buffers; D002712:Chlorides; D002586:Cesium; D011092:Polyethylene Glycols; D008156:Luciferases; C028019:cesium chloride