Enhanced cellular secretion of AAV2 by expression of foreign viral envelope proteins

Melissa A. Kotterman, James T. Koerber, Jung Soo Nam, Young Hoo Cho, Seung Hyun Kim, Jae-Hyung Jang, Kwang Il Lim

Research output: Contribution to journalArticle

Abstract

Recombinant adeno-associated virus (rAAV), a non-enveloped virus, is widely used in gene therapy clinical trials because it does not cause human disease, transduces both dividing and non-dividing cells, and mediates stable transgene expression for years in post-mitotic tissue. Extension of clinical use of rAAV is, however, considerably hampered by difficulties involved in large-scale production of the virus particles. For several serotypes of rAAV these difficulties often arise from the fact that assembled virus particles mainly stay inside of packaging cells, inevitably requiring lysis of cells to harvest virus particles and consequentially complicating downstream purification processes. Here, we show that introduction of foreign viral envelope protein genes, encoding for either VSVG or rabiesG, into packaging cells can remarkably enhance cellular secretion of rAAV2, the AAV serotype most often used in clinical applications. In the presence of the foreign genes, up to 49% of transducing rAAV2 particles were secreted. However, such great enhancement was not observed for rAAV3. Our experimental tests with exosome inhibitors indicated that VSVG-mediated cellular secretion of rAAV2, unlike rabiesG-mediated one, heavily relies upon cellular pathways involving exosomes. Ultimately, an improved understanding of rAAV secretion mechanisms may simplify the production and purification processes for large-scale batches of the virus.

Original languageEnglish
Pages (from-to)108-114
Number of pages7
JournalBiochemical Engineering Journal
Volume93
DOIs
Publication statusPublished - 2015 Jan 5

Fingerprint

Viral Envelope Proteins
Dependovirus
Viruses
Virion
Exosomes
Proteins
Product Packaging
Transgenes
Genetic Therapy
Genes
Purification
Packaging
Clinical Trials
Gene therapy
Gene encoding
Tissue
Serogroup

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Environmental Engineering
  • Biomedical Engineering

Cite this

Kotterman, Melissa A. ; Koerber, James T. ; Nam, Jung Soo ; Cho, Young Hoo ; Kim, Seung Hyun ; Jang, Jae-Hyung ; Lim, Kwang Il. / Enhanced cellular secretion of AAV2 by expression of foreign viral envelope proteins. In: Biochemical Engineering Journal. 2015 ; Vol. 93. pp. 108-114.
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Enhanced cellular secretion of AAV2 by expression of foreign viral envelope proteins. / Kotterman, Melissa A.; Koerber, James T.; Nam, Jung Soo; Cho, Young Hoo; Kim, Seung Hyun; Jang, Jae-Hyung; Lim, Kwang Il.

In: Biochemical Engineering Journal, Vol. 93, 05.01.2015, p. 108-114.

Research output: Contribution to journalArticle

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