Abstract
Recombinant viral vectors have been developed for use as therapeutic agents and for the introduction of exogenous genes into living cells. However, little is known about the viability and stability of such recombinant viruses during storage, transport and delivery under various conditions. We describe here an analysis of the stability of an adenoviral vector in crude solutions of cell lysates during freezing and thawing and during storage at various temperatures in the presence and in the absence of glycerol. For example, the titer of adenoviruses in crude lysates of infected cells was reduced only ten‐fold or three‐fold after two hundred rounds of freezing and thawing or after incubation at 28°C for 14 days, respectively. Our observations indicate that recombinant adenoviral vector was more stable than expected both during freezing and thawing and during storage at low temperatures. Our results confirm the importance of appropriate conditions for the delivery and transport of recombinant adenoviral vectors.
Keywords: Recombinant adenovirus, Banking, Gene therapy
Full Text
The Full Text of this article is available as a PDF (3.4 MB).
http://www.nih.gov/news/panelrep.html. Orkin, S. and Motulsky, A. G. Report and recommendation of the panel to assess the NIH investment in research in gene therapy.
REFERENCES
- 1. ) Yang , E. Y. , Flake , A. W. and Adzick , N. S.Prospects for fetal gene therapy . Semin. Perinatol. , 23 , 524 – 534 ( 1999. ). [DOI] [PubMed] [Google Scholar]
- 2. ) Anderson , W. F.Gene therapy. The best of times, the worst of times . Science , 288 , 627 – 629 ( 2000. ). [DOI] [PubMed] [Google Scholar]
- 3. ) Williams , D. A. and Smith , F. O.Progress in the use of gene transfer methods to treat genetic blood diseases . Hum. Gene Ther. , 11 , 2059 – 2066 ( 2000. ). [DOI] [PubMed] [Google Scholar]
- 4. ) Hitt , M. , Addison , C. and Graham , F.Human adenovirus vectors for gene transfer into mammalian cells . Adv. Pharmacol. , 40 , 137 – 206 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 5. ) Nyberg‐Hoffman , C. and Aguilar‐Cordova , E.Instability of adenoviral vectors during transport and its implication for clinical studies . Nat. Med. , 5 , 955 – 957 ( 1999. ). [DOI] [PubMed] [Google Scholar]
- 6. ) Niwa , H. , Yamamura , K. and Miyazaki , J.Efficient selection for high‐expression transfectants with a novel eukaryotic vector . Gene , 108 , 193 – 199 ( 1991. ). [DOI] [PubMed] [Google Scholar]
- 7. ) Kanegae , Y. , Lee , G. , Sato , Y. , Tanaka , M. , Nakai , M. , Sakaki , T. and Saito , I.Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site‐specific Cre recombinase . Nucleic Acids Res. , 23 , 3816 – 3821 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8. ) Nyberg‐Hoffman , C. , Shabram , P. , Li , W. , Giroux , D. and Aguilar‐Cordova , E.Sensitivity and reproducibility in adenoviral infectious titer determination . Nat. Med. , 4 , 808 – 811 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 9. ) Graham , F. L. , Smiley , J. , Russell , W. C. and Nairn , R.Characteristics of a human cell line transformed by DNA from human adenovirus type 5 . J. Gen. Virol. , 36 , 59 – 74 ( 1977. ). [DOI] [PubMed] [Google Scholar]
- 10. ) Precious , B. and Russell , W. C.Growth, purification and titration of adenoviruses . In “ Virology: a Practical Approach ,” ed. Mahy B. W. J. , pp. 193 – 205 ( 1985. ). IRL Press; , Oxford . [Google Scholar]
- 11. ) Yoshida , Y. and Hamada , H.Adenovirus‐mediated indueible gene expression through tetracycline‐controllable transactivator with nuclear localization signal . Biochem. Biophys. Res. Commun. , 230 , 426 – 430 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 12. ) Kanegae , Y. , Makimura , M. and Saito , I.A simple and efficient method for purification of infectious recombinant adenovirus . Jpn. J. Med. Sci. Biol. , 47 , 157 – 166 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 13. ) Croyle , M. A. , Yu , Q.‐C. and Wilson , J. M.Development of a rapid method for the PEGylation of adenoviruses with enhanced transduction and improved stability under harsh storage conditions . Hum. Gene Ther. , 11 , 1713 – 1722 ( 2000. ). [DOI] [PubMed] [Google Scholar]
