PMID- 26015972
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20150527
LR  - 20200930
IS  - 2329-0501 (Print)
IS  - 2329-0501 (Electronic)
IS  - 2329-0501 (Linking)
VI  - 1
DP  - 2014
TI  - Different protein composition and functional properties of adeno-associated 
      virus-6 vector manufactured from the culture medium and cell lysates.
PG  - 14031
LID - 10.1038/mtm.2014.31 [doi]
AB  - Vectors based on recombinant adeno-associated viruses (rAAV) attract a growing 
      interest for human gene therapy. Recently, it was shown that many rAAV serotypes 
      produced by transient transfection of human embryonic kidney 293 cell line 
      (HEK293) are efficiently released into culture medium and functionally equivalent 
      to those purified from cell lysates. Here, we report that HEK293 cells produce 
      and secrete Galectin 3-binding protein (huG3BP), a protein that efficiently binds 
      rAAV6 in vivo. Importantly, intracellular G3BP and secreted G3BP have different 
      properties: while the secreted protein had the same electrophoretic mobility as 
      serum huG3BP and interacted with rAAV6, intracellular protein migrated faster and 
      did not bind rAAV6. Consequently, rAAV6 purified from culture medium (secreted, 
      rAAV6-S) was physically associated with huG3BP while rAAV6 harvested from cell 
      lysates (cellular, rAAV6-C) was huG3BP-free. After systemic injections, rAAV6-S 
      bound to huG3BP was 3 times less efficient compared to rAAV6-C and induced an 
      immune response against huG3BP protein. Our findings show that protein content of 
      rAAVs purified from culture medium or from cell lysates can be different and 
      these differences may impact vector efficacy and/or immune response.
FAU - Denard, Jerome
AU  - Denard J
AD  - Biomarkers Department, Genethon, 1 bis rue de l'Internationale , Evry, France.
FAU - Jenny, Christine
AU  - Jenny C
AD  - Biomarkers Department, Genethon, 1 bis rue de l'Internationale , Evry, France.
FAU - Blouin, Veronique
AU  - Blouin V
AD  - Universite de Nantes-Inserm UMR649 , Nantes, France.
FAU - Moullier, Philippe
AU  - Moullier P
AD  - Universite de Nantes-Inserm UMR649 , Nantes, France ; Molecular Genetics and 
      Microbiology Department, University of Florida , Gainesville, Florida, USA.
FAU - Svinartchouk, Fedor
AU  - Svinartchouk F
AD  - Biomarkers Department, Genethon, 1 bis rue de l'Internationale , Evry, France.
LA  - eng
PT  - Journal Article
DEP - 20140730
PL  - United States
TA  - Mol Ther Methods Clin Dev
JT  - Molecular therapy. Methods & clinical development
JID - 101624857
PMC - PMC4362353
EDAT- 2014/01/01 00:00
MHDA- 2014/01/01 00:01
PMCR- 2014/07/30
CRDT- 2015/05/28 06:00
PHST- 2014/03/20 00:00 [received]
PHST- 2014/05/19 00:00 [revised]
PHST- 2014/06/05 00:00 [accepted]
PHST- 2015/05/28 06:00 [entrez]
PHST- 2014/01/01 00:00 [pubmed]
PHST- 2014/01/01 00:01 [medline]
PHST- 2014/07/30 00:00 [pmc-release]
AID - S2329-0501(16)30098-5 [pii]
AID - 10.1038/mtm.2014.31 [doi]
PST - epublish
SO  - Mol Ther Methods Clin Dev. 2014 Jul 30;1:14031. doi: 10.1038/mtm.2014.31. 
      eCollection 2014.