Isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column.
Hum Gene Ther
; 12(1): 71-6, 2001 Jan 01.
Article
em En
| MEDLINE
| ID: mdl-11177544
ABSTRACT
One of the most promising gene transfer vectors in human clinical trials is AAV2. The quality of the vector preparations is a key element in obtaining reliable and reproducible data in preclinical studies. However, established protocols either result in impure, low infectious virus (CsCl2 gradient centrifugation) or demand a high level of manual and technical skills (CsCl2 gradient centrifugation, iodixanol/heparin or HPLC purification). In this study, we present an easy-to-do single-step column purification (SSCP) of AAV2 by gravity flow based on its affinity to heparin, without ultracentrifugation. Various vector preparations generated by our method reproducibly showed high titers, infectivity, and purity. In vivo, our single-step column-purified AAV2 vectors mediate significantly higher transduction efficiency compared with conventional protocols. Investigators still unsatisfied with previously published techniques or new to the field of AAV production may find in our method an interesting alternative.
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Base de dados:
MEDLINE
Assunto principal:
Terapia Genética
/
Dependovirus
/
Vetores Genéticos
Tipo de estudo:
Guideline
/
Risk_factors_studies
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Hum Gene Ther
Assunto da revista:
GENETICA MEDICA
/
TERAPEUTICA
Ano de publicação:
2001
Tipo de documento:
Article
País de afiliação:
Estados Unidos