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The DOI matches its Crossref record; 6 of the 6 references it cites were found in a registry or an index.

High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency

Ayuso, E, Mingozzi, F, Montane, J, Leon, X, Anguela, X M, Haurigot, V, Edmonson, S A, Africa, L, Zhou, S, High, K A, Bosch, F, Wright, J F·2010·Gene Therapy
DOI 10.1038/gt.2009.157
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Abstract

The purity of adeno-associated virus (AAV) vector preparations has important implications for both safety and efficacy of clinical gene transfer. Early-stage screening of candidates for AAV-based therapeutics ideally requires a purification method that is flexible and also provides vectors comparable in purity and potency to the prospective investigational product manufactured for clinical studies. The use of cesium chloride (CsCl) gradient-based protocols provides the flexibility for purification of different serotypes; however, a commonly used first-generation CsCl-based protocol was found to result in AAV vectors containing large amounts of protein and DNA impurities and low transduction efficiency in vitro and in vivo. Here, we describe and characterize an optimized, second-generation CsCl protocol that incorporates differential precipitation of AAV particles by polyethylene glycol, resulting in higher yield and markedly higher vector purity that correlated with better transduction efficiency observed with several AAV serotypes in multiple tissues and species. Vectors purified by the optimized CsCl protocol were found to be comparable in purity and functional activity to those prepared by more scalable, but less flexible serotype-specific purification processes developed for manufacture of clinical vectors, and are therefore ideally suited for pre-clinical studies supporting translational research.

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Crossref · 10.1038/gt.2009.157

Cites

6 of 6 references verified0 citation errors · 0 not checkable

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  • Steady-state levels of lanosterol, lathosterol, and desmosterol were reduced in the striata of R6/2-Tom mice compared with wt-Tom mice.

    “As expected, the steady-state levels of all cholesterol precursors were reduced in the striata of R6/2-Tom mice compared with wt-Tom mice (Fig. 2H-J) 19,20,27,28 .”

    From SREBP2 gene therapy targeting striatal astrocytes ameliorates Huntington’s disease phenotypes (2021) · cites this paper as [28]

  • All three doses of cholesterol infused directly into the striata of R6/2 mice through osmotic mini-pumps prevented cognitive defects, while only the highest dose attenuated motor phenotypes.

    “Indeed, we demonstrated that all three doses prevented cognitive defects, while only the highest dose attenuated motor phenotypes 28 .”

    From SREBP2 gene therapy targeting striatal astrocytes ameliorates Huntington’s disease phenotypes (2021) · cites this paper as [28]

  • Enhanced cholesterol biosynthesis may participate in inducing clearance pathways.

    “At the neuropathological level, increased hSREBP2 in the striatum of R6/2 mice reduced the number and size of muHTT aggregates, supporting the notion that enhanced cholesterol biosynthesis may participate in inducing clearance pathways, as we recently showed in the striata of HD mice following striatal infusion of cholesterol through osmotic mini-pumps 28 .”

    From SREBP2 gene therapy targeting striatal astrocytes ameliorates Huntington’s disease phenotypes (2021) · cites this paper as [28]

  • Similar data were obtained upon cholesterol delivery to the brain of this mouse model via brain-permeable polymeric nanoparticles or mini-pumps.

    “Similar data were obtained upon cholesterol delivery to the brain of this mouse model via brain-permeable polymeric nanoparticles or mini-pumps 27,28 .”

    From SREBP2 gene therapy targeting striatal astrocytes ameliorates Huntington’s disease phenotypes (2021) · cites this paper as [28]

Citations

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Research works printed but not linked: 23

  1. Clark KR, Liu X, McGrath JP, Johnson PR . Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. Hum Gene Ther 1999; 10: 1031–1039. · DOI
  2. Davidoff AM, Ng CY, Sleep S, Gray J, Azam S, Zhao Y et al. Purification of recombinant adeno-associated virus type 8 vectors by ion exchange chromatography generates clinical grade vector stock. J Virol Methods 2004; 121: 209–215. · DOI
  3. Gao G, Qu G, Burnham MS, Huang J, Chirmule N, Joshi B et al. Purification of recombinant adeno-associated virus vectors by column chromatography and its performance in vivo. Hum Gene Ther 2000; 11: 2079–2091. · DOI
  4. Kaludov N, Handelman B, Chiorini JA . Scalable purification of adeno-associated virus type 2, 4, or 5 using ion-exchange chromatography. Hum Gene Ther 2002; 13: 1235–1243. · DOI
  5. Qu G, Bahr-Davidson J, Prado J, Tai A, Cataniag F, McDonnell J et al. Separation of adeno-associated virus type 2 empty particles from genome containing vectors by anion-exchange column chromatography. J Virol Methods 2007; 140: 183–192. · DOI
  6. Smith RH, Ding C, Kotin RM . Serum-free production and column purification of adeno-associated virus type 5. J Virol Methods 2003; 114: 115–124. · DOI
  7. Xiao X, Li J, Samulski RJ . Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus. J Virol 1998; 72: 2224–2232. · DOI · Paper page
  8. Gao GP, Alvira MR, Wang L, Calcedo R, Johnston J, Wilson JM . Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc Natl Acad Sci USA 2002; 99: 11854–11859. · DOI · Paper page
  9. Mingozzi F, Meulenberg JJ, Hui DJ, Basner-Tschakarjan E, Hasbrouck NC, Edmonson SA et al. AAV-1-mediated gene transfer to skeletal muscle in humans results in dose-dependent activation of capsid-specific T cells. Blood 2009; 114: 2077–2086. · DOI · Paper page
  10. Stroes ES, Nierman MC, Meulenberg JJ, Franssen R, Twisk J, Henny CP et al. Intramuscular administration of AAV1-lipoprotein lipase S447X lowers triglycerides in lipoprotein lipase-deficient patients. Arterioscler Thromb Vasc Biol 2008; 28: 2303–2304. · DOI · Paper page
  11. Cideciyan AV, Aleman TS, Boye SL, Schwartz SB, Kaushal S, Roman AJ et al. Human gene therapy for RPE65 isomerase deficiency activates the retinoid cycle of vision but with slow rod kinetics. Proc Natl Acad Sci USA 2008; 105: 15112–15117. · DOI
  12. Kaplitt MG, Feigin A, Tang C, Fitzsimons HL, Mattis P, Lawlor PA et al. Safety and tolerability of gene therapy with an adeno-associated virus (AAV) borne GAD gene for Parkinson's disease: an open label, phase I trial. Lancet 2007; 369: 2097–2105. · DOI · Paper page
  13. Grieger JC, Choi VW, Samulski RJ . Production and characterization of adeno-associated viral vectors. Nat Protoc 2006; 1: 1412–1428. · DOI
  14. Vandenberghe L, Lock M, Xiao R, Sandhu A, Lin J, Korn M et al. Efficient serotype-dependent release of functional vector into the supernatant during AAV manufacturing. Mol Ther 2009; 17: S271.
  15. Blankinship MJ, Gregorevic P, Chamberlain JS . Gene therapy strategies for Duchenne muscular dystrophy utilizing recombinant adeno-associated virus vectors. Mol Ther 2006; 13: 241–249. · DOI
  16. Brantly ML, Spencer LT, Humphries M, Conlon TJ, Spencer CT, Poirier A et al. Phase I trial of intramuscular injection of a recombinant adeno-associated virus serotype 2 alphal-antitrypsin (AAT) vector in AAT-deficient adults. Hum Gene Ther 2006; 17: 1177–1186. · DOI
  17. Mas A, Montane J, Anguela XM, Munoz S, Douar AM, Riu E et al. Reversal of type 1 diabetes by engineering a glucose sensor in skeletal muscle. Diabetes 2006; 55: 1546–1553. · DOI
  18. Margaritis P, Roy E, Aljamali MN, Downey HD, Giger U, Zhou S et al. Successful treatment of canine hemophilia by continuous expression of canine FVIIa. Blood 2009; 113: 3682–3689. · DOI
  19. Nathwani AC, Gray JT, McIntosh J, Ng CY, Zhou J, Spence Y et al. Safe and efficient transduction of the liver after peripheral vein infusion of self-complementary AAV vector results in stable therapeutic expression of human FIX in nonhuman primates. Blood 2007; 109: 1414–1421. · DOI
  20. Zhong L, Li B, Mah CS, Govindasamy L, Agbandje-McKenna M, Cooper M et al. Next generation of adeno-associated virus 2 vectors: point mutations in tyrosines lead to high-efficiency transduction at lower doses. Proc Natl Acad Sci USA 2008; 105: 7827–7832. · DOI · Paper page
  21. Matsushita T, Elliger S, Elliger C, Podsakoff G, Villarreal L, Kurtzman GJ et al. Adeno-associated virus vectors can be efficiently produced without helper virus. Gene Therapy 1998; 5: 938–945. · DOI
  22. Sommer JM, Smith PH, Parthasarathy S, Isaacs J, Vijay S, Kieran J et al. Quantification of adeno-associated virus particles and empty capsids by optical density measurement. Mol Ther 2003; 7: 122–128. · DOI · Paper page
  23. Herzog RW, Hagstrom JN, Kung SH, Tai SJ, Wilson JM, Fisher KJ et al. Stable gene transfer and expression of human blood coagulation factor IX after intramuscular injection of recombinant adeno-associated virus. Proc Natl Acad Sci USA 1997; 94: 5804–5809. · DOI

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Metadata

Authors
Ayuso, E., Mingozzi, F., Montane, J., Leon, X., Anguela, X. M., Haurigot, V, Edmonson, S. A., Africa, L., Zhou, S., High, K. A., Bosch, F., & Wright, J. F.
Venue
Gene Therapy
Publisher
Springer Science and Business Media LLC
Published
2009-12-03
Volume
17
Issue
4
Pages
503-510
ISSN
  • 0969-7128
  • 1476-5462
DOI
10.1038/gt.2009.157
Cited by
272
References
6
Language
en
Licence

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