Rapid and simplified purification of recombinant adeno-associated virus.

Journal of Virological Methods
Ping GuoGeorge K Gittes

Abstract

Preclinical gene therapy studies both in vitro and in vivo require high purity preparations of adeno-associated virus (AAV). Current methods for purification of AAV entail the use of centrifugation over either a CsCl or iodixanol gradient, or the use of chromatography. These methods can be cumbersome and expensive, necessitating ultrahigh speed gradient centrifugation or, for chromatography the use of other expensive equipment. In addition, these methods are time consuming, and the viral yield is not high. Currently no commercial purification kits are available for other than AAV serotype 2. A simplified method was used for the purification of AAV, with a viral yield that is able to be used effectively in adult and embryo mice. The method does not require ultrahigh speed gradient centrifugation nor chromatography. Instead, polyethylene glycol (PEG)/aqueous two-phase partitioning is used to remove soluble proteins from the PEG8000 precipitated virus-protein mixture. The procedure obtained rapidly up to 95% recovery of high quality purified AAV. The entire purification process, including HEK293 cell transfection, can be completed readily within one week, with purity seemingly higher than that obtained after one round of CsCl grad...Continue Reading

References

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Citations

Feb 19, 2014·Diabetologia·Xiangwei XiaoGeorge K Gittes
Jan 19, 2016·Frontiers in Molecular Neuroscience·Mu Qiao, Joshua R Sanes
Jan 7, 2016·Proceedings of the National Academy of Sciences of the United States of America·Ken BerglundUte Hochgeschwender
Feb 14, 2015·Biotechnology and Bioengineering·Piergiuseppe NestolaManuel J T Carrondo
Feb 1, 2015·Journal of Chromatography. a·Christopher Ladd EffioJürgen Hubbuch
Mar 19, 2014·Proceedings of the National Academy of Sciences of the United States of America·Xiangwei XiaoGeorge K Gittes
Dec 14, 2017·Scientific Reports·Mairobys SocorroFarzad Esni
Oct 9, 2013·Laboratory Investigation; a Journal of Technical Methods and Pathology·Ping GuoGeorge K Gittes
Apr 15, 2018·Journal of Lipid Research·Xin Hui Derryn ChanShigeki Sugii
Sep 2, 2017·Journal of Neuroscience Research·Sung Young ParkUte Hochgeschwender
Apr 23, 2020·Preparative Biochemistry & Biotechnology·Neha N Gharat, Virendra K Rathod
Sep 21, 2019·Scientific Reports·Toyokazu KimuraMarkus Michael Bachschmid
Jan 8, 2013·Journal of Nippon Medical School = Nippon Ika Daigaku Zasshi·Koichi MiyakeYukihiko Hirai
Feb 15, 2018·PeerJ·Geert CremersHuub J M Op den Camp
Nov 20, 2019·International Journal of Molecular Sciences·Rebecca C FeinerKristian M Müller
Apr 9, 2019·Journal of Neuroscience Research·Ken BerglundRobert E Gross
Mar 18, 2021·Neuroscience Letters·Akinsola Raphael AkinyemiQiang Liu
Aug 19, 2020·Journal of Neuroscience Methods·Ayumu Konno, Hirokazu Hirai
May 21, 2021·The EMBO Journal·Jungjoo ParkWon-Suk Chung

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