Enabling development of commercial-ready lentiviral vector manufacturing processes using stable producer cell lines

2021 ◽  
Vol 7 (9) ◽  
pp. 981-993
Author(s):  
Peter Archibald ◽  
Anthony Shillings
Keyword(s):  
Cell Lines ◽  
Lentiviral Vector ◽  
Manufacturing Processes ◽  
Producer Cell

scholarly journals Efficient construction of producer cell lines for a SIN lentiviral vector for SCID-X1 gene therapy by concatemeric array transfection

Blood ◽  
2009 ◽  
Vol 113 (21) ◽  
pp. 5104-5110 ◽  
Author(s):  
Robert E. Throm ◽  
Annastasia A. Ouma ◽  
Sheng Zhou ◽  
Anantharaman Chandrasekaran ◽  
Timothy Lockey ◽  
...  
Keyword(s):  
Cell Lines ◽  
Large Scale ◽  
Fluorescent Protein ◽  
Lentiviral Vector ◽  
Cell Clone ◽  
Severe Combined Immunodeficiency ◽  
Interleukin 2 ◽  
Retroviral Vectors ◽  
Scale Production ◽  
Producer Cell

AbstractRetroviral vectors containing internal promoters, chromatin insulators, and self-inactivating (SIN) long terminal repeats (LTRs) may have significantly reduced genotoxicity relative to the conventional retroviral vectors used in recent, otherwise successful clinical trials. Large-scale production of such vectors is problematic, however, as the introduction of SIN vectors into packaging cells cannot be accomplished with the traditional method of viral transduction. We have derived a set of packaging cell lines for HIV-based lentiviral vectors and developed a novel concatemeric array transfection technique for the introduction of SIN vector genomes devoid of enhancer and promoter sequences in the LTR. We used this method to derive a producer cell clone for a SIN lentiviral vector expressing green fluorescent protein, which when grown in a bioreactor generated more than 20 L of supernatant with titers above 107 transducing units (TU) per milliliter. Further refinement of our technique enabled the rapid generation of whole populations of stably transformed cells that produced similar titers. Finally, we describe the construction of an insulated, SIN lentiviral vector encoding the human interleukin 2 receptor common γ chain (IL2RG) gene and the efficient derivation of cloned producer cells that generate supernatants with titers greater than 5 × 107 TU/mL and that are suitable for use in a clinical trial for X-linked severe combined immunodeficiency (SCID-X1).

scholarly journals Development of inducible EIAV-based lentiviral vector packaging and producer cell lines

Gene Therapy ◽  
2009 ◽  
Vol 16 (6) ◽  
pp. 805-814 ◽  
Author(s):  
H J Stewart ◽  
M A Leroux-Carlucci ◽  
C J M Sion ◽  
K A Mitrophanous ◽  
P A Radcliffe
Keyword(s):  
Cell Lines ◽  
Lentiviral Vector ◽  
Producer Cell

scholarly journals Scalable Lentiviral Vector Production Using Stable HEK293SF Producer Cell Lines

2017 ◽  
Vol 28 (6) ◽  
pp. 330-339 ◽  
Author(s):  
Aziza P. Manceur ◽  
Howard Kim ◽  
Vanja Misic ◽  
Nadejda Andreev ◽  
July Dorion-Thibaudeau ◽  
...  
Keyword(s):  
Cell Lines ◽  
Lentiviral Vector ◽  
Producer Cell

scholarly journals A strategy to obtain recombinant cell lines with high expression levels. Lentiviral vector-mediated transgenesis

2011 ◽  
Vol 5 (S8) ◽  
Author(s):  
Claudio Prieto ◽  
Diego Fontana ◽  
Marina Etcheverrigaray ◽  
Ricardo Kratje
Keyword(s):  
Cell Lines ◽  
Lentiviral Vector ◽  
High Expression ◽  
Expression Levels ◽  
Recombinant Cell

A bicistronic SIN-lentiviral vector containing G156A MGMT allows selection and metabolic correction of hematopoietic protoporphyric cell lines

2003 ◽  
Vol 5 (9) ◽  
pp. 737-747 ◽  
Author(s):  
Emmanuel Richard ◽  
Fabien Géronimi ◽  
Magalie Lalanne ◽  
Cécile Ged ◽  
Isabelle Redonnet-Vernhet ◽  
...  
Keyword(s):  
Cell Lines ◽  
Lentiviral Vector ◽  
Metabolic Correction

[23] Recombinant adeno-associated viral vector production using stable packaging and producer cell lines

2002 ◽  
pp. 393-413 ◽  
Author(s):  
Lydia C. Mathews ◽  
John T. Gray ◽  
Mark R. Gallagher ◽  
Richard O. Snyder
Keyword(s):  
Cell Lines ◽  
Viral Vector ◽  
Producer Cell

scholarly journals Packaging Cells Based on Inducible Gene Amplification for the Production of Adeno-Associated Virus Vectors

1998 ◽  
Vol 72 (9) ◽  
pp. 7024-7031 ◽  
Author(s):  
Naoki Inoue ◽  
David W. Russell
Keyword(s):  
Cell Lines ◽  
Gene Amplification ◽  
Large Scale ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Scale Production ◽  
Adeno Associated Virus ◽  
Tetracycline Transactivator ◽  
Producer Cell

ABSTRACT Although vectors based on adeno-associated virus (AAV) offer several unique advantages, their usage has been hampered by the difficulties encountered in vector production. In this report, we describe a new AAV packaging system based on inducible amplification of integrated helper and vector constructs containing the simian virus 40 (SV40) replication origin. The packaging and producer cell lines developed express SV40 T antigen under the control of the reverse tetracycline transactivator system, which allows inducible amplification of chromosomal loci linked to the SV40 origin. Culturing these cells in the presence of doxycycline followed by adenovirus infection resulted in helper and vector gene amplification as well as higher vector titers. Clonal producer cell lines generated vector titers that were 10 times higher than those obtained by standard methods, with approximately 104vector particles produced per cell. These stocks were free of detectable replication-competent virus. The lack of a transfection step combined with the reproducibility of stable producer lines makes this packaging method ideally suited for the large-scale production of vector stocks for human gene therapy.

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