PMID- 30073177
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20201001
IS  - 2329-0501 (Print)
IS  - 2329-0501 (Electronic)
IS  - 2329-0501 (Linking)
VI  - 10
DP  - 2018 Sep 21
TI  - Helper-free Production of Laboratory Grade AAV and Purification by Iodixanol 
      Density Gradient Centrifugation.
PG  - 1-7
LID - 10.1016/j.omtm.2018.05.001 [doi]
AB  - Adeno-associated virus (AAV) is one of the most promising gene therapy vectors 
      and is widely used as a gene delivery vehicle for basic research. As AAV 
      continues to become the vector of choice, it is increasingly important for new 
      researchers to have access to a simplified production and purification protocol 
      for laboratory grade recombinant AAV. Here we report a detailed protocol for 
      serotype independent production of AAV using a helper-free HEK293 cell system 
      followed by iodixanol gradient purification, a method described earlier.(1) While 
      the core principals of this mammalian AAV production system are unchanged, there 
      have been significant advancements in the production and purification procedure 
      that serve to boost yield, maximize efficiency, and increase the purity of AAV 
      preps. Using this protocol, we are able to constantly obtain high quantities of 
      laboratory grade AAV particles (>5 x 10(12) vg) in a week's time, largely 
      independent of serotype.
FAU - Crosson, Sean M
AU  - Crosson SM
AD  - Department of Pediatrics, Division of Cell and Molecular Therapy, University of 
      Florida, Gainesille, FL, USA.
FAU - Dib, Peter
AU  - Dib P
AD  - Department of Anatomy and Cell Biology, University of Florida, Gainesille, FL, 
      USA.
FAU - Smith, J Kennon
AU  - Smith JK
AD  - Department of Biochemistry and Molecular Biology, University of Florida, 
      Gainesille, FL, USA.
FAU - Zolotukhin, Sergei
AU  - Zolotukhin S
AD  - Department of Pediatrics, Division of Cell and Molecular Therapy, University of 
      Florida, Gainesille, FL, USA.
LA  - eng
GR  - F31 DC015751/DC/NIDCD NIH HHS/United States
GR  - R01 DC012819/DC/NIDCD NIH HHS/United States
PT  - Journal Article
DEP - 20180508
PL  - United States
TA  - Mol Ther Methods Clin Dev
JT  - Molecular therapy. Methods & clinical development
JID - 101624857
PMC - PMC6069679
EDAT- 2018/08/04 06:00
MHDA- 2018/08/04 06:01
PMCR- 2018/05/08
CRDT- 2018/08/04 06:00
PHST- 2018/02/12 00:00 [received]
PHST- 2018/05/01 00:00 [accepted]
PHST- 2018/08/04 06:00 [entrez]
PHST- 2018/08/04 06:00 [pubmed]
PHST- 2018/08/04 06:01 [medline]
PHST- 2018/05/08 00:00 [pmc-release]
AID - S2329-0501(18)30047-0 [pii]
AID - 10.1016/j.omtm.2018.05.001 [doi]
PST - epublish
SO  - Mol Ther Methods Clin Dev. 2018 May 8;10:1-7. doi: 10.1016/j.omtm.2018.05.001. 
      eCollection 2018 Sep 21.