Scalable production of adeno-associated virus type 2 vectors via suspension transfection

Biotechnology and Bioengineering
Joon Young ParkEui-Cheol Jo

Abstract

Vectors derived from adeno-associated virus type 2 (AAV2) are promising gene delivery vehicles, but it is still challenging to get the large number of recombinant adeno-associated virus (rAAV) particles required for large animal and clinical studies. Current transfection technology requires adherent cultures of HEK 293 cells that can only be expanded by preparing multiple culture plates. A single large-scale suspension culture could replace these multiple culture preparations, but there is currently no effective co-transfection scheme for generating rAAV from cells in suspension culture. Here, we weaned HEK 293 cells to suspension culture using hydrogel-coated six-well culture plates and established an efficient transfection strategy suitable for these cells. Then the cultures were gradually scaled up. We used linear polyethylenimine (PEI) to mediate transfection and obtained high transfection efficiencies ranging from 54% to 99%, thereby allowing efficient generation of rAAV vectors. Up to 10(13) rAAV particles and, more importantly, up to 10(11) infectious particles were generated from a 2-L bioreactor culture. The suspension-transfection strategy of this study facilitates the homogeneous preparation of rAAV at a large scale,...Continue Reading

References

Aug 1, 1995·Proceedings of the National Academy of Sciences of the United States of America·O BoussifJ P Behr
Apr 29, 1998·Human Gene Therapy·A SalvettiP Moullier
Mar 30, 2000·The Journal of Gene Medicine·R O Snyder
Aug 10, 2000·Molecular Therapy : the Journal of the American Society of Gene Therapy·J M AllenA D Miller
Oct 6, 2000·Molecular Therapy : the Journal of the American Society of Gene Therapy·X LiuK R Clark
Mar 28, 2001·The Journal of Gene Medicine·M K ChuahT VandenDriessche
Mar 28, 2001·The Journal of Gene Medicine·L DrittantiM Vega
Apr 25, 2001·The Journal of Gene Medicine·A KichlerO Danos
Jan 12, 2002·Molecular Therapy : the Journal of the American Society of Gene Therapy·Sylvia BrunnerErnst Wagner
Aug 23, 2002·Proceedings of the National Academy of Sciences of the United States of America·Guang-Ping GaoJames M Wilson
Nov 5, 2002·Methods : a Companion to Methods in Enzymology·Dirk Grimm
Nov 5, 2002·Methods : a Companion to Methods in Enzymology·Sergei ZolotukhinRichard O Snyder
Nov 13, 2002·Human Gene Therapy·Masashi UrabeRobert M Kotin
Nov 20, 2002·Molecular Therapy : the Journal of the American Society of Gene Therapy·Nicole BrumentAnna Salvetti
Dec 3, 2002·Current Opinion in Biotechnology·Richard O Snyder, Terence R Flotte
Jun 6, 2003·Molecular Therapy : the Journal of the American Society of Gene Therapy·Dirk GrimmJuergen A Kleinschmidt
Nov 20, 2003·Journal of Virological Methods·Richard H SmithRobert M Kotin
Aug 3, 2004·Biotechnology and Bioengineering·Madiha DerouaziFlorian M Wurm
May 27, 2005·Human Gene Therapy·S Zolotukhin
Apr 23, 2008·Current Protocols in Human Genetics·A D Miller
Nov 13, 2008·Cytotechnology·Philippe GirardFlorian M Wurm
Nov 13, 2008·Cytotechnology·E J Schlaeger, K Christensen

❮ Previous
Next ❯

Citations

Sep 18, 2010·Bioprocess and Biosystems Engineering·Olivier HenryAmine Kamen
Nov 16, 2001·Trends in Cardiovascular Medicine·C May, M Sadelain
Nov 14, 2000·Molecular Medicine Today·P E Monahan, R J Samulski
Jan 30, 2002·Trends in Molecular Medicine·Ji-Won Yoon, Hee-Sook Jun
Apr 5, 2008·Molecular Therapy : the Journal of the American Society of Gene Therapy·Haifeng Chen
May 3, 2011·Human Molecular Genetics·Robert M Kotin
Jun 16, 2010·Journal of Bioscience and Bioengineering·Yu-Kuo LiuLie-Ding Shiau
Oct 7, 2015·Molecular Therapy : the Journal of the American Society of Gene Therapy·Joshua C GriegerRichard Jude Samulski
Nov 1, 2015·Human Molecular Genetics·Johannes C M van der Loo, J Fraser Wright
Jan 4, 2018·Critical Reviews in Biotechnology·Sonia Gutiérrez-GranadosFrancesc Gòdia
Sep 29, 2019·Cytotechnology·Peifeng ChenFrank Arnold
Sep 26, 2020·Biotechnology Journal·Jihad El Andari, Dirk Grimm
Jul 17, 2020·Molecular Therapy. Methods & Clinical Development·Huiren ZhaoW Hans Meisen
May 7, 2021·Biotechnology Advances·Sha ShaRichard D Braatz

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cell-Type Specific Viral Vectors

Viral vectors are used in biological research and therapy to deliver genetic material into cells. However, the efficiency of viral vectors varies depending on the cell type. Here is the latest research on cell-type-specific viral vectors.

Cell-Type-Specific Viral Vectors (ASM)

Viral vectors are used in biological research and therapy to deliver genetic material into cells. However, the efficiency of viral vectors varies depending on the cell type. Here is the latest research on cell-type-specific viral vectors.

Cell-Type-Specific Viral Vectors

Viral vectors are used in biological research and therapy to deliver genetic material into cells. However, the efficiency of viral vectors varies depending on the cell type. Here is the latest research on cell-type-specific viral vectors.