Ultrafiltered recombinant AAV8 vector can be safely administered in vivo and efficiently transduces liver

Mark D. Kleven, Michelle M. Gomes, Aaron M. Wortham, Caroline Enns, Christoph Kahl

Research output: Contribution to journalArticle

Abstract

Viral vectors are extensively purified for use in biomedical research, in order to separate biologically active virus particles and to eliminate production related impurities that are assumed to be detrimental to the host. For recombinant adeno-associated virus (rAAV) vectors this is typically accomplished using density gradient-based methods, which are tedious and require specialized ultracentrifugation equipment. In order to streamline the preparation of rAAV vectors for pilot and small animal studies, we recently devised a simple ultrafiltration approach that permits rapid virus concentration and partial removal of production-related impurities. Here we show that systemic administration of such rapidly prepared (RP) rAAV8 vectors in mice is safe and efficiently transduces the liver. Across a range of doses, delivery of RP rAAV8-CMV-eGFP vector induced enhanced green fluorescent protein (eGFP) expression in liver that was comparable to that obtained from a conventional iodixanol gradient- purified (IP) vector. Surprisingly, no liver inflammation or systemic cytokine induction was detected in RP rAAV injected animals, revealing that residual impurities in the viral vector preparation are not deleterious to the host. Together, these data demonstrate that partially purified rAAV vector can be safely and effectively administered in vivo. The speed and versatility of the RP method and lack of need for cumbersome density gradients or expensive ultracentrifuge equipment will enable more widespread use of RP prepared rAAV vectors, such as for pilot liver gene transfer studies.

Original languageEnglish (US)
Article numbere0194728
JournalPLoS One
Volume13
Issue number4
DOIs
StatePublished - Apr 1 2018

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Dependovirus
Liver
Viruses
liver
green fluorescent protein
Equipment and Supplies
ultracentrifugation
Impurities
Ultracentrifugation
biomedical research
Ultrafiltration
ultrafiltration
virion
gene transfer
Virion
Animals
Biomedical Research
animals
cytokines
protein synthesis

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Ultrafiltered recombinant AAV8 vector can be safely administered in vivo and efficiently transduces liver. / Kleven, Mark D.; Gomes, Michelle M.; Wortham, Aaron M.; Enns, Caroline; Kahl, Christoph.

In: PLoS One, Vol. 13, No. 4, e0194728, 01.04.2018.

Research output: Contribution to journalArticle

Kleven, Mark D. ; Gomes, Michelle M. ; Wortham, Aaron M. ; Enns, Caroline ; Kahl, Christoph. / Ultrafiltered recombinant AAV8 vector can be safely administered in vivo and efficiently transduces liver. In: PLoS One. 2018 ; Vol. 13, No. 4.
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